Recombinant Anti-alpha Tubulin acetyl K40 antibody [EPR16772] - Rat IgG2a (Chimeric - BSA and Azide free) (ab289872)

This data was developed using ab289863, the same antibody clone in a different buffer formulation. 

Blocking and diluting buffer and concentration: 

Exposure time: Lane 1-6: 3.25 seconds Lane 7-8: 5.5 seconds

This data was developed using ab289863, the same antibody clone in a different buffer formulation. 

Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling alpha Tubulin (acetyl K40) with ab289863 at 1/100 followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection kit). Positive staining on human cerebrum. The section was incubated with ab289863 for 30 mins at room temperature and followed by specific rat IgG antibody (ab102248) for 8 mins, immunostaining was performed on a Leica Biosystems BOND^® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control: PBS was used instead of primary antibody followed by a ready to use secondary antibody LeicaDS9800 (BOND™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

This data was developed using ab289863, the same antibody clone in a different buffer formulation. 

Immunohistochemical analysis of paraffin-embedded rat cerebrum tissue labeling alpha Tubulin (acetyl K40) with ab289863 at 1/500 followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection kit). Positive staining on rat cerebrum. The section was incubated with ab289863 for 30 mins at room temperature and followed by specific rat IgG antibody (ab102248) for 8 minutes, immunostaining was performed on a Leica Biosystems BOND^® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control: PBS was used instead of primary antibody followed by a ready to use secondary antibody LeicaDS9800 (BOND™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

This data was developed using ab289863, the same antibody clone in a different buffer formulation. 

Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling alpha Tubulin (acetyl K40) with ab289863 at 1/500 followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection kit). Positive staining on mouse cerebrum. The section was incubated with ab289863 for 30 mins at room temperature and followed by specific rat IgG antibody (ab102248) for 8 mins, immunostaining was performed on a Leica Biosystems BOND^® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control: PBS was used instead of primary antibody followed by a ready to use secondary antibody LeicaDS9800 (BOND™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

This data was developed using ab289863, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa cells labeling alpha Tubulin (acetyl K40) with ab289863 at 1/100 dilution (10.81 µg/ml), followed by ab96971 Goat Anti-Rat IgG Fc (DyLight^® 488) preadsorbed antibody at 1/500 dilution (Green). Confocal image showing increased cytoplasmic staining in HeLa cells treated with Trichostatin A (500 ng/ml) for 4 hours. ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control: PBS was used instead of primary antibody followed by secondary antibody is ab96971 Goat Anti-Rat IgG Fc (DyLight^® 488) preadsorbed at 1/500 dilution.

This data was developed using ab289863, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 cells labeling alpha Tubulin (acetyl K40) with ab289863 at 1/100 dilution (10.81 µg/ml), followed by ab96971 Goat Anti-Rat IgG Fc (DyLight^® 488) preadsorbed antibody at 1/500 dilution (Green). Confocal image showing increased cytoplasmic staining in NIH/3T3 cells treated with Trichostatin A (500 ng/ml) for 4 hours. ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control: PBS was used instead of primary antibody followed by secondary antibody is ab96971 Goat Anti-Rat IgG Fc (DyLight^® 488) preadsorbed at 1/500 dilution.

This data was developed using ab289863, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized C6 cells labeling alpha Tubulin (acetyl K40) with ab289863 at 1/100 dilution (10.81 µg/ml), followed by ab96971 Goat Anti-Rat IgG Fc (DyLight^® 488) preadsorbed antibody at 1/500 dilution (Green). Confocal image showing increased cytoplasmic staining in C6 cells treated with Trichostatin A (500 ng/ml) for 4 hours. ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control: PBS was used instead of primary antibody followed by secondary antibody is ab96971 Goat Anti-Rat IgG Fc (DyLight^® 488) preadsorbed at 1/500 dilution.

This data was developed using ab289863, the same antibody clone in a different buffer formulation.

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) treated with 500 ng/ml Trichostatin A for 4 hours (Red) / Untreated control (Green) cells labeling alpha Tubulin (acetyl K40) with ab289863 at 1/1000 dilution (Red) compared with a Rat monoclonal IgG (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rat IgG Fc (DyLight 488, ab96971) at 1/2000 dilution was used as the secondary antibody.