Anti-6X His tag® 抗体 [EPR20547] - ChIP Grade

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (AB213204)

Immunohistochemical analysis of paraffin-embedded human liver tissue labeling 6X His tag® with ab213204 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Negative control : No staining on human liver.

Counter stained with Hematoxylin.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (AB213204)

Immunohistochemical analysis of agarose-embedded HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with a His-tagged Staphylococcus aureus cas9 (J7RUA5; aa1-1053; 125kDa) construct labeling 6X His tag® with ab213204 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Left image : Positive staining on HEK-293T transfected with a His-tagged Staphylococcus aureus cas9 (J7RUA5; aa1-1053; 125kDa) construct. Right image : No staining on HEK-293T transfected with an empty expression vector.

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (AB213204)

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with GFP-Myc-His vector labeling 6X His tag® with ab213204 (right panel) at 1/5000 dilution compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (left panel). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) (ab150079) at 1/2000 dilution was used as the secondary antibody.

Gate is set between transfected and untransfected HEK-293T cells.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (AB213204)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cells transfected with GFP-Myc-His vector expression construct labeling 6X His tag® with ab213204 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (white). Confocal image showing positive staining on HEK-293T cells transfected with GFP-Myc-His vector expression construct.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) (red) at 1/200 dilution.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

• ChIP

Supplier Data

ChIP - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (AB213204)

Chromatin was prepared from MCF7 (human breast adenocarcinoma cell line) cells transfected with 6X His-tagged GATA3 according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25μg of chromatin, 2μg of ab213204 (blue), and 20μl of A/G sepharose beads slurry (10μl of sepharose A beads + 10μl of sepharose G beads). 2μg of rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).

ChIP was performed according to the literature (PMID : 22951069).

• IP

Supplier Data

Immunoprecipitation - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (AB213204)

His-tagged Staphylococcus aureus cas9 was immunoprecipitated from 0.35 mg of HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with a His-tagged Staphylococcus aureus cas9 (J7RUA5; aa1-1053; 125 kDa) construct, whole cell lysate with ab213204 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab213204 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution

Lane 1 : HEK-293T transfected with His-tagged Staphylococcus aureus cas9 construct, whole cell lysate 10 μg (Input).

Lane 2 : ab213204 IP in HEK-293T transfected with His-tagged Staphylococcus aureus cas9 construct, whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab213204 in HEK-293T transfected with His-tagged Staphylococcus aureus cas9 construct, whole cell lysate.

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : 1 second.

All lanes:

Immunoprecipitation - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

Observed band size: 125 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (AB213204)

Immunohistochemical analysis of paraffin-embedded rat stomach tissue labeling 6X His tag® with ab213204 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Negative control : No staining on rat stomach.

Counter stained with Hematoxylin.

• WB

Lab

Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (AB213204)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

This antibody does not cross-react with human KCNQ1, KCNQ2, KCNQ4 or KCNQ5 by western blot.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa); Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000)(96kDa).

All lanes:

Western blot - Anti-KCNQ3 antibody [EPR25237-111] (<a href='/products/primary-antibodies/kcnq3-antibody-epr25237-111-ab325554'>ab325554</a>) at 1/1000 dilution

Lane 1:

293T cells transfected with an empty vector containing a his-tag, whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a human KCNQ1 expression vector containing a his-tag, whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a human KCNQ2 expression vector containing a his-tag, whole cell lysate at 4 µg

Lane 4:

293T cells transfected with a human KCNQ3 expression vector containing a his-tag, whole cell lysate at 4 µg

Lane 5:

293T cells transfected with a human KCNQ4 expression vector containing a his-tag, whole cell lysate at 20 µg

Lane 6:

293T cells transfected with a human KCNQ5 expression vector containing a his-tag, whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 96 kDa,36 kDa

false

Exposure time: 6s

• WB

Lab

Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (AB213204)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000) (23KDa); Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-TSLP antibody [EPR30575-508] (<a href='/products/primary-antibodies/tslp-antibody-epr30575-508-ab327005'>ab327005</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a myc-His-tag® whole cell lysate at 5 µg

Lane 2:

293T cells transfected with a human TSLP expression vector containing a myc-His-tag® whole cell lysate at 5 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 23 kDa,36 kDa

false

Exposure time: 15s

• WB

Lab

Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (AB213204)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

This antibody does not cross-react with full-length human Kallikrein-6, Kallikrein-13, Kallikrein-2 and Kallikrein-8.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000)(30-37KDa).

All lanes:

Western blot - Anti-Kallikrein-11 antibody [EPR31011-525R] (<a href='/products/primary-antibodies/kallikrein-11-antibody-epr31011-525r-ab326566'>ab326566</a>) at 1/1000 dilution

Lane 1:

His-tagged full-length human Kallikrein-11 recombinant protein at 10 ng

Lane 2:

His-tagged full-length human Kallikrein-6 recombinant protein at 10 ng

Lane 3:

His-tagged full-length human Kallikrein-13 recombinant protein at 10 ng

Lane 4:

His-tagged full-length human Kallikrein-2 recombinant protein at 10 ng

Lane 5:

His-tagged full-length human Kallikrein-8 recombinant protein at 10 ng

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 37 kDa,30-37 kDa

false

Exposure time: 70s

• WB

Supplier Data

Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (AB213204)

Amongst human GGT homologous proteins, this antibody reacts with GGT1 and GGT3 but not GGT2.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-GGT1/GGT + GGT3 antibody [EPR28715-82] (<a href='/products/primary-antibodies/ggt1-ggt-ggt3-antibody-epr28715-82-ab316768'>ab316768</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a myc-His-tag®, whole cell lysate 7 at 7 µg

Lane 2:

293T cells transfected with a human GGT1 expression vector containing a myc-His-tag®, whole cell lysate at 7 µg

Lane 3:

293T cells transfected with a human GGT2 expression vector containing a myc-His-tag®, whole cell lysate at 7 µg

Lane 4:

293T cells transfected with a human GGT3 expression vector containing a myc-His-tag®, whole cell lysate at 7 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 60-75 kDa,36 kDa,60 kDa,75 kDa

false

Exposure time: 37s

• WB

Lab

Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (AB213204)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

This antibody does not cross-react with overexpressed human NR4A1 or NR4A3 by western blot.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000) (45-67KDa); Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-Nurr1 antibody [EPR26338-73] (<a href='/products/primary-antibodies/nurr1-antibody-epr26338-73-ab327018'>ab327018</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a myc-His-tag® whole cell lysate at 10 µg

Lane 2:

293T cells transfected with a human Nurr1 expression vector containing a myc-His-tag® whole cell lysate at 10 µg

Lane 3:

293T cells transfected with a human NR4A3 expression vector containing a myc-His-tag® whole cell lysate at 10 µg

Lane 4:

293T cells transfected with a human NR4A1 expression vector containing a myc-His-tag® whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 45-67 kDa,36 kDa

false

Exposure time: 26s

• WB

Supplier Data

Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (AB213204)

This antibody does not cross-react with human STAG2.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-SA1 antibody [EPR26227-76] (<a href='/products/primary-antibodies/sa1-antibody-epr26227-76-ab317301'>ab317301</a>) at 1/1000 dilution

Lane 1:

His-tagged Human STAG1 protein

Lane 2:

His-tagged Human STAG2 protein

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 19 kDa,38 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (AB213204)

This antibody recognizes overexpressed human ATAD3A isoform 2 and isoform 1 (UniProt ID : Q9NVI7).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-ATAD3A antibody [EPR27484-26] (<a href='/products/primary-antibodies/atad3a-antibody-epr27484-26-ab317252'>ab317252</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) transfected with an empty vector containing a myc-His-tag®, whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a Human ATAD3A isoform 2 expression vector containing a myc-His-tag®, whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a Human ATAD3A isoform 1 expression vector containing a myc-His-tag®, whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 66 kDa,76 kDa,36 kDa

false

Exposure time: 62s

• WB

Supplier Data

Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (AB213204)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The antibody detects Human STAT4 (phospho Y693).

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

In Western blot, anti-STAT4 antibody (ab284408) staining at 1/1000 dilution.

In Western blot, anti-His antibody (ab213204) staining at 1/5000 dilution.

In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution.

All lanes:

Western blot - Anti-STAT4 (phospho Y693) antibody [EPR27081-49] (<a href='/products/primary-antibodies/stat4-phospho-y693-antibody-epr27081-49-ab313630'>ab313630</a>) at 1/1000 dilution

Lane 1:

293T cells transfected with a Human STAT4 expression vector containing a His-tag were treated with treated with 100 mM Calyculin A for 30 minutes whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a Human STAT4 (Y693F mutation) expression vector containing a His-tag were treated with 100 mM Calyculin A for 30 minutes whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 85 kDa

true

Exposure time: 180s

• WB

Lab

Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (AB213204)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

This antibody does not cross-react with human CXCL2 and CXCL3.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa);Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000)(11KDa).

All lanes:

Western blot - Anti-CXCL1 antibody [EPR31264-520] (<a href='/products/primary-antibodies/cxcl1-antibody-epr31264-520-ab326942'>ab326942</a>) at 1/1000 dilution

Lane 1:

293T cells transfected with an empty vector containing a his-tag, whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a human CXCL1 expression vector containing a his-tag, whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a human CXCL2 expression vector containing a his-tag, whole cell lysate at 20 µg

Lane 4:

293T cells transfected with a human CXCL3 expression vector containing a his-tag, whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/1000 dilution

Observed band size: 11 kDa,36 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (AB213204)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.
Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) was used at 1/1,000,000 dilution

All lanes:

Western blot - Anti-PTBP1 + PTBP2 + PTBP3 antibody [EPR9890] (<a href='/products/primary-antibodies/ptbp2-antibody-epr9890-ab154787'>ab154787</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) + OE-empty whole cell lysate at 20 µg

Lane 2:

293T + OE-PTBP2 whole cell lysate at 20 µg

Lane 3:

293T + OE-PTBP1 whole cell lysate at 20 µg

Lane 4:

293T + OE-PTBP3 whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/20000 dilution

Predicted band size: 55 kDa

false

Exposure time: 3s

• WB

Lab

Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (AB213204)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

This antibody does not cross-react with overexpressed human SCD1 by western blot.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000)(38, 41KDa);Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-SCD5 antibody [MJF-D29765-175] (<a href='/products/primary-antibodies/scd5-antibody-mjf-d29765-175-ab327055'>ab327055</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a myc-His-tag® whole cell lysate at 5 µg

Lane 2:

293T cells transfected with a human SCD5 expression vector containing a myc-His-tag® whole cell lysate at 5 µg

Lane 3:

293T cells transfected with a human SCD1 expression vector containing a myc-His-tag® whole cell lysate at 5 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 38 kDa,41 kDa,36 kDa

false

Exposure time: 8s

• WB

Lab

Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (AB213204)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa); Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000)(53KDa).

All lanes:

Western blot - Anti-PNPLA3 I148M variant antibody [EPR30478-584] (<a href='/products/primary-antibodies/pnpla3-i148m-variant-antibody-epr30478-584-ab325644'>ab325644</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a myc-His-tag®, whole cell lysate at 20 µg

Lane 2:

293T transfected with a human PNPLA3(I148M mutation) expression vector containing a myc-His-tag® whole cell lysate at 20 µg

Lane 3:

293T transfected with a human PNPLA3 (WT) expression vector containing a myc-His-tag® whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 52 kDa,36 kDa,53 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (AB213204)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000) (83KDa); Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-Hormone sensitive lipase/HSL (phospho S660) antibody [EPR30648-593] (<a href='/products/primary-antibodies/hormone-sensitive-lipase-hsl-phospho-s660-antibody-epr30648-593-ab326331'>ab326331</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a myc-His-tag® whole cell lysate (untreated membrane) at 5 µg

Lane 2:

293T cells transfected with a human wild-type HSL expression vector containing a myc-His-tag® whole cell lysate (untreated membrane) at 5 µg

Lane 3:

293T cells transfected with a human wild-type HSL expression vector containing a myc-His-tag®, treated with 100nM Calyculin A for 30 minutes whole cell lysate (untreated membrane) at 5 µg

Lanes 4 and 8:

293T cells transfected with a human HSL (mutation S650A) expression vector containing a myc-His-tag®, treated with 100nM Calyculin A for 30 minutes whole cell lysate (alkaline phosphatase treated membrane) at 5 µg

Lane 5:

293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a myc-His-tag® whole cell lysate (alkaline phosphatase treated membrane) at 5 µg

Lane 6:

293T cells transfected with a human wild-type HSL expression vector containing a myc-His-tag® whole cell lysate (alkaline phosphatase treated membrane) at 5 µg

Lane 7:

293T cells transfected with a human wild-type HSL expression vector containing a myc-His-tag®, treated with 100nM Calyculin A for 30 minutes whole cell lysate (alkaline phosphatase treated membrane) at 5 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 83 kDa,36 kDa

false

Exposure time: 6s

• WB

Lab

Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (AB213204)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

This antibody does not cross-react with human MURF3 and MURF2.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000)(37-100KDa).

All lanes:

Western blot - Anti-MURF1 antibody [EPR29784-48] (ab327341) at 1/1000 dilution

Lane 1:

Untagged human MURF1 fragment at 10 ng

Lane 2:

His-tagged human MURF3 fragment at 10 ng

Lane 3:

His-tagged human MURF2 fragment at 10 ng

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 16 kDa,37-100 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (AB213204)

Blocking buffer and dilution concentration : 5% NFDM/TBST.

Lanes 1 - 4:

Western blot - Anti-TrkA (phospho Y674)+TrkB (phospho Y706)+TrkC (phospho Y709) antibody [EPR22298-67] (<a href='/products/primary-antibodies/trka-phospho-y674trkb-phospho-y706trkc-phospho-y709-antibody-epr22298-67-ab229908'>ab229908</a>) at 1/1000 dilution

Lanes 1 - 4:

Western blot - Anti-TrkA (pY674) + TrkB (pY706) + TrkC (pY709) antibody [EPR22298-67] - BSA and Azide free (<a href='/products/primary-antibodies/trka-phospho-y674trkb-phospho-y706trkc-phospho-y709-antibody-epr22298-67-bsa-and-azide-free-ab264070'>ab264070</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a His-tag, whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a human TrkA expression vector containing a His-tag, whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a human TrkB expression vector containing a His-tag, whole cell lysate at 20 µg

Lane 4:

293T cells transfected with a human TrkC expression vector containing a His-tag, whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 140 kDa

false

Exposure time: 15s

• WB

Lab

Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (AB213204)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

This antibody does not cross-react with overexpressed human DLL3 or DLL4 by western blot.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000)(85, 30KDa); Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-DLL1 antibody [EPR29655-26] (ab327088) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a myc-His-tag® whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a human DLL1 expression vector containing a myc-His-tag® whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a human DLL3 expression vector containing a myc-His-tag® whole cell lysate at 20 µg

Lane 4:

293T cells transfected with a human DLL4 expression vector containing a myc-His-tag® whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 85 kDa,30 kDa,36 kDa

false

Exposure time: 8s

• WB

Lab

Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (AB213204)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

This antibody does not cross-react with overexpressed human CPLX2 by western blot.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa); Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000) (17KDa).

All lanes:

Western blot - Anti-CPLX1 antibody [EPR31022-42] (ab327002) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a myc-His-tag® whole cell lysate at 10 µg

Lane 2:

293T cells transfected with a human CPLX1 expression vector containing a myc-His-tag® whole cell lysate at 10 µg

Lane 3:

293T cells transfected with a human CPLX2 expression vector containing a myc-His-tag® whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 17 kDa,36 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (AB213204)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

This antibody does not cross-react with human TRAK2.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa);Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000)(130KDa).

All lanes:

Western blot - Anti-TRAK1 antibody [EPR30574-567] (<a href='/products/primary-antibodies/trak1-antibody-epr30574-567-ab327353'>ab327353</a>) at 1/1000 dilution

Lane 1:

293T cells transfected with an empty vector containing a his-tag, whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a human TRAK1 expression vector containing a his-tag, whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a human TRAK2 expression vector containing a his-tag, whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 130 kDa,36 kDa

false

Exposure time: 8s

• WB

Supplier Data

Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (AB213204)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

UV-C treatment induces phosphorylation of Chk1 at Ser345 (PMID : 20609246).

The identity of the bands higher than 150 kDa are unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

In Western blot, Anti-Chk1 antibody [EP691Y] (ab40866) staining at 56KDa dilution.

All lanes:

Western blot - Anti-Chk1 (phospho S345) antibody [EPR30270-566] (<a href='/products/primary-antibodies/chk1-phospho-s345-antibody-epr30270-566-ab325675'>ab325675</a>) at 1/1000 dilution

Lane 1:

Untreated Synchronized G1/S HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate (untreated membrane) at 20 µg

Lane 2:

Synchronized G1/S HeLa treated with 100 J/m2 UV-C, then recovery for 1 hour whole cell lysate (untreated membrane) at 20 µg

Lane 3:

Untreated Synchronized G1/S HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate (phosphatase treated membrane) at 20 µg

Lane 4:

Synchronized G1/S HeLa treated with 100 J/m2 UV-C, then recovery for 1 hour whole cell lysate (phosphatase treated membrane) at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 56 kDa,36 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (AB213204)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

This antibody does not cross-react with recombinant human VPS4B by western blot.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000)(48KDa).

All lanes:

Western blot - Anti-VPS4a antibody [EPR30587-24] (<a href='/products/primary-antibodies/vps4a-antibody-epr30587-24-ab327386'>ab327386</a>) at 1/1000 dilution

Lane 1:

His-tagged human VPS4A recombinant protein at 10 ng

Lane 2:

His-tagged human VPS4B recombinant protein at 10 ng

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 48 kDa

false

Exposure time: 48s

• WB

Lab

Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (AB213204)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa); Anti-6X His tag antibody [EPR20547]-CHIP Grade (ab213204) (1 : 5000)(35-50KDa).

All lanes:

Western blot - Anti-Cyclin E1(Phospho T77) antibody [EPR28889-10] (ab327426) at 1/5000 dilution

Lane 1:

293T cells transfected with a human wild-type CCNE1 expression vector containing a His-tag, whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a human CCNE1 (T77A) expression vector containing a His-tag, whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/200000 dilution

Observed band size: 35-50 kDa,36 kDa

false

Exposure time: 15s

• WB

Lab

Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (AB213204)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The antibody shows weak reactivity with HEK293T cell lines overexpressing human Ret (Y1062A mutant)

In Western blot, Anti-Ret antibody (ab134100) (1 : 1000) (150KDa); Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000)(150KDa); Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) 1 : 100000 (15Kda).

All lanes:

Western blot - Anti-Ret (phospho Y1062) antibody [EPR30639-507] (<a href='/products/primary-antibodies/ret-phospho-y1062-antibody-epr30639-507-ab326572'>ab326572</a>) at 1/1000 dilution

Lane 1:

293T cells transfected with an empty vector containing a his-tag treated with 100uM pervanadate for 10 min whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a human wild-type Ret expression vector containing a his-tag were treated with 100uM Pervanadate for 10 mins, whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a human Ret(Y1062A mutant) expression vector containing a his-tag were treated with 100uM Pervanadate for 10 mins, whole cell lysate at 2 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 150 kDa,15 kDa

false

Exposure time: 8s

• WB

Supplier Data

Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (AB213204)

This antibody can recognize human SHANK1, SHANK2 and SHANK3. .

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-SHANK1 + SHANK2 + SHANK3 antibody [EPR26549-87] (<a href='/products/primary-antibodies/shank1-shank2-shank3-antibody-epr26549-87-ab317608'>ab317608</a>) at 1/1000 dilution

Lane 1:

His-tagged human SHANK2 recombinant fragment at 10 ng

Lane 2:

His-tagged human SHANK1 recombinant fragment at 10 ng

Lane 3:

His-tagged human SHANK3 recombinant fragment at 10 ng

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 26 kDa

false

Exposure time: 125s

• WB

Supplier Data

Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (AB213204)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-Chk1 (phospho S345) antibody [EPR30270-566] (<a href='/products/primary-antibodies/chk1-phospho-s345-antibody-epr30270-566-ab325675'>ab325675</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) transfected with an empty vector containing a myc-His-tag® whole cell lysate at 20 µg

Lane 2:

293T transfected with a human wild-type Chk1 expression vector containing a myc-His-tag® whole cell lysate at 20 µg

Lane 3:

293T transfected with a human Chk1 (S345A mutation) expression vector containing a myc-His-tag® whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 56 kDa,36 kDa

false

Exposure time: 37s

• WB

Lab

Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (AB213204)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The antibody does not detect full-length recombinant JNK2 and JNK3 by Western Blot.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000)(52KDa).

All lanes:

Western blot - Anti-JNK3 antibody [EPR29664-135] (<a href='/products/primary-antibodies/jnk3-antibody-epr29664-135-ab326931'>ab326931</a>) at 1/1000 dilution

Lane 1:

His-tagged human JNK3 full-length recombinant protein at 10 ng

Lane 2:

His-tagged human JNK1 full-length recombinant protein at 10 ng

Lane 3:

His-tagged human JNK2 full-length recombinant protein at 10 ng

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 52 kDa

false

Exposure time: 37s

• WB

Lab

Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (AB213204)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The antibody reacts with both the ROR gamma and ROR gamma t isoforms.

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) 1 : 100000 (15Kda);
Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000)(37,55kDa).

All lanes:

Western blot - Anti-ROR gamma antibody [EPR27137-10] (<a href='/products/primary-antibodies/ror-gamma-antibody-epr27137-10-ab325651'>ab325651</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a myc-His-tag® whole cell lysate at 20 µg

Lane 2:

293T cells transfected with human ROR gamma t expression vector containing a myc-His-tag® whole cell lysate at 20 µg

Lane 3:

293T cells transfected with human ROR gamma expression vector containing a myc-His-tag® whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 37 kDa,55 kDa,15 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (AB213204)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

This antibody does not cross-react with human MAPK8IP1, MAPK8IP2 or MAPK8IP4.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000)(240KDa).

All lanes:

Western blot - Anti-JIP3 antibody [EPR30425-79] (<a href='/products/primary-antibodies/jip3-antibody-epr30425-79-ab326560'>ab326560</a>) at 1/1000 dilution

Lane 1:

293T cells transfected with a human MAPK8IP3 expression vector containing a his-tag, whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a human MAPK8IP1 expression vector containing a his-tag, whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a human MAPK8IP2 expression vector containing a his-tag, whole cell lysate at 20 µg

Lane 4:

293T cells transfected with a human MAPK8IP4 expression vector containing a his-tag, whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 240 kDa

false

Exposure time: 3s

• WB

Lab

Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (AB213204)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, Anti-Histone H3 antibody [EPR16987] - Loading Control (ab176842) (1 : 100000) (36KDa); Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000)(150-250kDa).

All lanes:

Western blot - Anti-SOS1 (phospho Y1196) antibody [TF7-28-23] (<a href='/products/primary-antibodies/sos1-phospho-y1196-antibody-tf7-28-23-ab325591'>ab325591</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) transfected with an empty vector containing a myc-His-tag® whole cell lysate at 20 µg

Lane 2:

293T transfected with a human wild-type SOS1 expression vector containing a myc-His-tag® whole cell lysate at 20 µg

Lane 3:

293T transfected with a human SOS1 (Y1196A mutation) expression vector containing a myc-His-tag® whole cell lysate at 20 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Observed band size: 152 kDa,36 kDa,150-250 kDa

true

Exposure time: 180s

• WB

Lab

Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (AB213204)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

This antibody does not cross-react with human CTSK, CTSS or CTSV.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000)(30-75KDa).

All lanes:

Western blot - Anti-Cathepsin L/MEP antibody [EPR30887-518] (<a href='/products/primary-antibodies/cathepsin-l-mep-antibody-epr30887-518-ab326362'>ab326362</a>) at 1/1000 dilution

Lane 1:

No tagged human CTSL recombinant fragment at 10 ng

Lane 2:

His-tagged human CTSK recombinant fragment at 10 ng

Lane 3:

His-tagged human CTSS recombinant fragment at 10 ng

Lane 4:

His-tagged human CTSV recombinant fragment at 10 ng

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 75 kDa,30-75 kDa

false

Exposure time: 48s

• WB

Lab

Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (AB213204)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

This antibody does not cross-react with recombinant human SORCS3 by western blot.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa); Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000) (130KDa).

All lanes:

Western blot - Anti-SorCS1 antibody [EPR30536-522] (<a href='/products/primary-antibodies/sorcs1-antibody-epr30536-522-ab325598'>ab325598</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) transfected with an empty vector containing a His-tag, whole cell lysate at 20 µg

Lane 2:

293T transfected with a human SORCS1 expression vector containing a His-tag, whole cell lysate at 20 µg

Lane 3:

293T transfected with a human SORCS3 expression vector containing a His-tag, whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 130 kDa,36 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (AB213204)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. Lysates were not boiled.

Lanes 1 - 2:

Western blot - Anti-VEGF Receptor 1 antibody [Y103] (<a href='/products/primary-antibodies/vegf-receptor-1-antibody-y103-ab32152'>ab32152</a>) at 1/1000 dilution

Lanes 1 - 2:

Western blot - Anti-VEGF Receptor 1 antibody [Y103] - BSA and Azide free (<a href='/products/primary-antibodies/vegf-receptor-1-antibody-y103-bsa-and-azide-free-ab210608'>ab210608</a>)

Lanes 1 - 2:

Western blot - Anti-VEGF Receptor 1 antibody [Y103] - Low endotoxin, Azide free (<a href='/products/primary-antibodies/vegf-receptor-1-antibody-y103-low-endotoxin-azide-free-ab184784'>ab184784</a>)

Lanes 1 - 2:

Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) at 1/5000 dilution

Lane 1:

HEK-293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a His tag whole cell lysate at 20 µg

Lane 2:

HEK-293T (human embryonic kidney epithelial cell) cells transfected with sFlt1 (P17948-2) expression vector containing a His -tag whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 77 kDa

false

Exposure time: 5s

• WB

Lab

Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (AB213204)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

This antibody does not cross-react with overexpressed human ZMIZ2 by western blot.

To minimize protein degradation, cells were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible. ZMIZ1 can undergo degradation easily.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa); Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000)(150KDa).

All lanes:

Western blot - Anti-Zinc finger MIZ domain-containing protein 1 antibody [EPR29807-525] (<a href='/products/primary-antibodies/zinc-finger-miz-domain-containing-protein-1-antibody-epr29807-525-ab326070'>ab326070</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a myc-His-tag® fresh whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a human ZMIZ2 expression vector containing a myc-His-tag® fresh whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a human ZMIZ1 expression vector containing a myc-His-tag® fresh whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 120 kDa,130 kDa,150 kDa,36 kDa

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (AB213204)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. This blot was developed using a high sensitivity ECL substrate. In Western blot, Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671) staining at 1/1000 dilution. Anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution.Anti-His antibody (ab213204) staining at 1/5000 dilution. Exposure time : 3 minutes.

All lanes:

Western blot - Anti-ATG16L1 (phospho S278) antibody [EPR19016] - Rat IgG1 (Chimeric) (<a href='/products/primary-antibodies/atg16l1-phospho-s278-antibody-epr19016-rat-igg1-chimeric-ab309495'>ab309495</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) transfected with an empty vector (vector control) containing a myc-His-tag whole cell lysate (untreated membrane) at 20 µg

Lane 2:

293T cells transfected with a human ATG16L1 expression vector containing a myc-His-tag whole cell lysate (untreated membrane) at 20 µg

Lane 3:

293T cells transfected with a human ATG16L1 (mutation S278A) expression vector containing a myc-His-tag whole cell lysate (untreated membrane) at 20 µg

Lane 4:

293T (human embryonic kidney epithelial cell) transfected with an empty vector (vector control) containing a myc-His-tag whole cell lysate (Alkaline phosphatase treated membrane) at 20 µg

Lane 5:

293T cells transfected with a human ATG16L1 expression vector containing a myc-His-tag whole cell lysate (Alkaline phosphatase treated membrane) at 20 µg

Lane 6:

293T cells transfected with a human ATG16L1 (mutation S278A) expression vector containing a myc-His-tag whole cell lysate (Alkaline phosphatase treated membrane) at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rat-igg-h-l-hrp-ab205720'>ab205720</a>) at 1/5000 dilution

Observed band size: 68 kDa

false