Anti-HA tag antibody [EPR22819-101] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HA tag antibody [EPR22819-101] - BSA and Azide free (AB256483)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-HA tag antibody [EPR22819-101] - BSA and Azide free (AB256483)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab236632).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-HA tag antibody [EPR22819-101] - BSA and Azide free (AB256483)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab236632).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HA tag antibody [EPR22819-101] - BSA and Azide free (AB256483)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.

• IP

Unknown

Immunoprecipitation - Anti-HA tag antibody [EPR22819-101] - BSA and Azide free (AB256483)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab236632).

All lanes:

Immunoprecipitation - Anti-HA tag antibody [EPR22819-101] (<a href='/en-us/products/primary-antibodies/ha-tag-antibody-epr22819-101-ab236632'>ab236632</a>)

Predicted band size: 64 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HA tag antibody [EPR22819-101] - BSA and Azide free (AB256483)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HA tag antibody [EPR22819-101] - BSA and Azide free (AB256483)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.