Anti-RYBP 抗体 [EPR13059(2)] - ChIP Grade

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (AB185971)

Immunohistochemical analysis of paraffin-embedded, Human placenta tissue labeling RYBP with ab185971 at a 1/100 dilution. Counter stained with hematoxylin.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (AB185971)

Intracellular Flow Cytometry analysis of paraformaldehyde-fixed K562 cells labeling RYBP with ab185971 at a 1/40 dilution and secondary antibody goat anti-rabbit IgG (FITC, red) at a 1/150 dilution, or negative control rabbit IgG (green).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (AB185971)

Immunofluorescence analysis of, paraformaldehyde-fixed, SW480 cells labeling RYBP with ab185971 at a 1/250 dilution. As secondary antibody goat anti-rabbit IgG (Alexa Fluor®555) was used at a 1/200. In blue DAPI staining.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (AB185971)

Intracellular Flow Cytometry analysis of A431 (Human epidermoid carcinoma epithelial cell) cells labeling RYBP with purified ab185971 at 1/20 dilution (11.7μg/mL) (Red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) (black). Unlabeled control - Unlabelled cells (blue).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (AB185971)

Immunofluorescence analysis of, paraformaldehyde-fixed, HepG2 cells labeling RYBP with ab185971 at a 1/250 dilution. As secondary antibody goat anti-rabbit IgG (Alexa Fluor®555) was used at a 1/200. In blue DAPI staining.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (AB185971)

Immunohistochemical analysis of paraffin-embedded, Human clear cell carcinoma kidney tissue labeling RYBP with ab185971 at a 1/100 dilution. Counter stained with hematoxylin.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IP

Lab

Immunoprecipitation - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (AB185971)

RYBP was immunoprecipitated from HEK-293 (human embryonic kidney epithelial cell) whole cell lysate 10 μg with ab185971 1/1000 dilution. Western blot was performed on the immunoprecipitate using ab185971 at 1/1000 dilution. Capture antibody, 1 : 30 dilution (2μg in 0.35mg lysates).

Rabbit monoclonal IgG (ab172730) Isotype Control was used instead of ab185971 in HEK-293 whole cell lysate (Lane 3).

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

To minimize protein degradation, cells were lysed immediately after harvest and then applied for Immunoprecipitation as soon as possible.

All lanes:

Immunoprecipitation - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971) at 1/1000 dilution

Lane 1:

HEK-293 (human embryonic kidney epithelial cell) whole cell lysate (Low Expression Sample) 10 μg

Lane 2:

HEK-293 (human embryonic kidney epithelial cell) whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 32 kDa

false

Exposure time: 15s

• ChIP

Unknown

ChIP - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (AB185971)

Chromatin was prepared from HEK293 cells according to the Abcam Dual X-ChIP protocol*. Cells were fixed with EGS for 30 minutes, then formaldehyde for 10 minutes.
The ChIP was performed with 25 µg of chromatin, 5 µg of ab185971 (red), and 20 µl of Protein A/G sepharose beads. 5 µg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
Primers and probes are located in the first kb of the transcribed region.
*http : //www.abcam.com/resources?keywords=X%20ChIP%20protocol

• IP

Lab

Immunoprecipitation - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (AB185971)

RYBP was immunoprecipitated from NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 10 μg with ab185971 1/1000 dilution. Western blot was performed on the immunoprecipitate using ab185971 at 1/1000 dilution. Capture antibody, 1 : 30 dilution (2μg in 0.35mg lysates).

Rabbit monoclonal IgG (ab172730) Isotype Control was used instead of ab185971 in NIH/3T3 whole cell lysate (Lane 3).

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

To minimize protein degradation, cells were lysed immediately after harvest and then applied for Immunoprecipitation as soon as possible.

All lanes:

Immunoprecipitation - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971) at 1/1000 dilution

Lane 1:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate (Low Expression Sample) 10 μg

Lane 2:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 32 kDa

false

Exposure time: 24s

• ChIP

Unknown

ChIP - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (AB185971)

Chromatin was prepared from NIH/3T3 cells according to the Abcam Dual X-ChIP protocol*. Cells were fixed with EGS for 30 minutes, then formaldehyde for 10 minutes.
The ChIP was performed with 25 µg of chromatin, 5 µg of ab185971 (red), and 20 µl of Protein A/G sepharose beads. 5 µg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
Primers and probes are located in the first kb of the transcribed region.
*http : //www.abcam.com/resources?keywords=X%20ChIP%20protocol

• WB

Supplier Data

Western blot - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (AB185971)

All lanes:

Western blot - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971) at 1/10000 dilution

All lanes:

HepG2 cell lysate at 20 µg

Secondary

All lanes:

goat anti-rabbit IgG, (H+L), peroxidase conjugated at 1/1000 dilution

Predicted band size: 25 kDa

Observed band size: 32 kDa

true

• WB

Supplier Data

Western blot - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (AB185971)

All lanes:

Western blot - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971) at 1/1000 dilution

Lane 1:

K562 cell lysate at 20 µg

Lane 2:

SW480 cell lysate at 20 µg

Secondary

All lanes:

goat anti-rabbit IgG, (H+L), peroxidase conjugated at 1/1000 dilution

Predicted band size: 25 kDa

Observed band size: 32 kDa

true

• WB

Supplier Data

Western blot - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (AB185971)

All lanes:

Western blot - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971) at 1/1000 dilution

All lanes:

rat PC12 cell lysate at 10 µg

Secondary

All lanes:

goat anti-rabbit IgG, (H+L), peroxidase conjugated at 1/1000 dilution

Predicted band size: 25 kDa

Observed band size: 32 kDa

true