Anti-HA tag 抗体 [EPR22819-101]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HA tag antibody [EPR22819-101] (AB236632)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human embryonic kidney epithelial cell) transfected with a HA Tag expression vector, (B) HEK-293T transfected with empty vector labelling HA tag with ab236632 at 1/5000 (0.124 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on HEK-293T transfected with a HA Tag expression vector (Image A) and no staining on (B) HEK-293T transfected with empty vector (Image B)
The section was incubated with ab236632 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND RX instrument.

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HA tag antibody [EPR22819-101] (AB236632)

Immunohistochemical analysis of paraffin-embedded human tonsil with ab236632 at 1/5000 dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on human tonsil.

The section was incubated with ab236632 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND RX instrument.

Counterstained with Hematoxylin.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-HA tag antibody [EPR22819-101] (AB236632)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293T (human embryonic kidney epithelial cell) transfected with HA-tagged TATA-box-binding protein (WT) expression vector labeling HA tag with ab236632 at 1/1000 (0.6 μg/ml) dilution, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000, (2 μg/ml) dilution (Green). Confocal image showing nuclear staining in HEK-293T cells transfected with HA-tagged TATA-box-binding protein (WT) expression vector is observed. Anti-HA.11 Epitope Tag antibody (Alexa Fluor^® 647) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 (2 μg/ml) dilution.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-HA tag antibody [EPR22819-101] (AB236632)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized HEK-293T (human embryonic kidney) transfected with TATA-box-binding protein (WT) expression vector containing HA-tag (Right) / HEK-293T (Left) cells labeling HA tag with ab236632 at 1/600 dilution. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

• IP

Unknown

Immunoprecipitation - Anti-HA tag antibody [EPR22819-101] (AB236632)

HA tag was immunoprecipitated from 0.35 mg HEK-293T (human embryonic kidney) transfected with TIM 4 (WT) expression vector containing a HA-GFP-tag and PDGFR TM domain whole cell lysate 10μg with ab236632 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab236632. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/2000 dilution.

Lane 1 : HEK-293T (human embryonic kidney) transfected with TIM 4 (WT) expression vector containing a HA-GFP-tag and PDGFR TM domain whole cell lysate 10μg.

Lane 2 : ab236632 IP in HEK-293T transfected with TIM 4 (WT) expression vector containing a HA-GFP-tag and PDGFR TM domain whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab236632 in HEK-293T transfected with TIM 4 (WT) expression vector containing a HA-GFP-tag and PDGFR TM domain whole cell lysate.

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

Exposure time : 3 seconds.

All lanes:

Immunoprecipitation - Anti-HA tag antibody [EPR22819-101] (ab236632)

Predicted band size: 64 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HA tag antibody [EPR22819-101] (AB236632)

Immunohistochemical analysis of paraffin-embedded Rat spleen with ab236632 at 1/5000 dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on Rat spleen.

The section was incubated with ab236632 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND RX instrument.

Counterstained with Hematoxylin.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HA tag antibody [EPR22819-101] (AB236632)

Immunohistochemical analysis of paraffin-embedded Mouse spleen with ab236632 at 1/5000 dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on Mouse spleen.

The section was incubated with ab236632 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND RX instrument.

Counterstained with Hematoxylin.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• WB

Supplier Data

Western blot - Anti-HA tag antibody [EPR22819-101] (AB236632)

Rfx Cas13d may cleave to two fragments close to C-term, and this antibody can recognize the full length and N-terminal fragment. HA tag antibody can recognize the full length and N-terminal fragment. His-tag antibody can recognize full length and C-terminal fragment because the His-tag of this overexpression vector located on C-term, HA-myc-tag of this overexpression vector located on N-term.

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

In Western blot, Anti-HA tag antibody [EPR22819-101] - (ab236632) staining at 1/1000 dilution.

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

All lanes:

Western blot - Anti-Rfx Cas13d (N-term) antibody [EPR23965-31] (<a href='/products/primary-antibodies/rfx-cas13d-n-term-antibody-epr23965-31-ab314741'>ab314741</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 with 5% NFDM/TBST

Lane 2:

293T cells transfected with a Ruminococcus flavefaciens cas13d expression vector containing a HA-myc-tag (N-term) and a His-tag® (C-term), whole cell lysate at 10 µg with 5% NFDM/TBST

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: 110 kDa,70 kDa,36 kDa,40 kDa

false

Exposure time: 6s

• WB

Lab

Western blot - Anti-HA tag antibody [EPR22819-101] (AB236632)

Antibodies used in the control panels are (Anti-TIM 4 antibody), ab9110 (Anti-HA tag antibody - ChIP Grade) and ab181602 (Anti-GAPDH antibody [EPR16891] - Loading Control).

Exposure time : 3 minutes.

Blocking and Diluting Buffer and concentration : 5% NFDM/TBST.

All lanes:

Western blot - Anti-HA tag antibody [EPR22819-101] (ab236632) at 1/1000 dilution

Lane 1:

HEK-293T (human embryonic kidney) transfected with an empty vector (vector control), containing a HA-myc-His-GFP-tag and PDGFR TM domain, whole cell lysate at 10 µg

Lane 2:

HEK-293T transfected with TIM 4 (WT) expression vector containing a HA-myc-His-GFP-tag and PDGFR TM domain, 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

Predicted band size: 64 kDa

Observed band size: 37 kDa,78 kDa

false