Anti-alpha Tubulin acetyl K40 antibody [EPR16772] - Rat IgG2a (Chimeric)

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin acetyl K40 antibody [EPR16772] - Rat IgG2a (Chimeric) (AB289863)

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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha Tubulin acetyl K40 antibody [EPR16772] - Rat IgG2a (Chimeric) (AB289863)

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.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-alpha Tubulin acetyl K40 antibody [EPR16772] - Rat IgG2a (Chimeric) (AB289863)

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.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin acetyl K40 antibody [EPR16772] - Rat IgG2a (Chimeric) (AB289863)

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.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin acetyl K40 antibody [EPR16772] - Rat IgG2a (Chimeric) (AB289863)

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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha Tubulin acetyl K40 antibody [EPR16772] - Rat IgG2a (Chimeric) (AB289863)

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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha Tubulin acetyl K40 antibody [EPR16772] - Rat IgG2a (Chimeric) (AB289863)

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.

• WB

Lab

Western blot - Anti-alpha Tubulin acetyl K40 antibody [EPR16772] - Rat IgG2a (Chimeric) (AB289863)

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

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

All lanes:

Western blot - Anti-alpha Tubulin acetyl K40 antibody [EPR16772] - Rat IgG2a (Chimeric) (ab289863) at 1/1000 dilution

Lane 1:

Untreated HeLa (Human epithelial cells from cervix adenocarcinoma), whole cell lysate at 20 µg

Lane 2:

HeLa treated with 500 ng/ml Trichostatin A for 4 hours, whole cell lysate at 20 µg

Lane 3:

Untreated NIH/3T3 (Mouse embyro fibroblast cells), whole cell lysate at 20 µg

Lane 4:

NIH/3T3 treated with 500 ng/ml Trichostatin A for 4 hours, whole cell lysate at 20 µg

Lane 5:

Untreated C6 (Rat glial tumor cells), whole cell lysate at 20 µg

Lane 6:

C6 treated with 500 ng/ml Trichostatin A for 4 hours, whole cell lysate at 20 µg

Lane 7:

Mouse brain tissue lysate at 20 µg

Lane 8:

Rat brain tissue lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 50 kDa

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