Anti-Histone H2B (mutated G53D) antibody [EPR26428-35] - BSA and Azide free

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Histone H2B (mutated G53D) antibody [EPR26428-35] - BSA and Azide free (AB305053)

This data was developed using ab305052, the same antibody clone in a different buffer formulation. Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HEK-293T (human embryonic kidney) cells transfected with a human Histone H2B expression vector containing a myc-his tag (Left) / HEK-293T transfected with a human Histone H2B (mutated G53D) expression vector containing a myc-his tag (Right) cells labelling Histone H2B (mutated G53D) with ab305052 at 1/500 dilution (0.1ug)/ Left and Right (Red) compared with a isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• ICC

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Immunocytochemistry - Anti-Histone H2B (mutated G53D) antibody [EPR26428-35] - BSA and Azide free (AB305053)

This data was developed using ab305052, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HEK-293T (human embryonic kidney epithelial cell) cells labelling Histone H2B (mutated G53D) with ab305052 at 1/500 (1.1 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing positive staining in HEK-293T cells transfected with a human H2BC4 (G54D) expression vector containing a myc tag and showing no staining in HEK-293T cells transfected with a human H2BC4 wildtype expression vector containing a myc tag. The image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. Myc-Tag Mouse mAb (Alexa Fluor® 647) was used to counterstain tubulin at 1/100 0.38 ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• ChIP

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ChIP - Anti-Histone H2B (mutated G53D) antibody [EPR26428-35] - BSA and Azide free (AB305053)

This data was developed using ab305052, the same antibody clone in a different buffer formulation.  Chromatin was prepared from 293T transfected with Histone H2B (mutated G53D) and 293T transfected with Histone H2B cells according to the Abcam X-ChIP protocol*. Cells were fixed with formaldehyde for 10min. The ChIP was performed with 25 µg of chromatin, 2 µg of ab305052 (red), or 2 µg of rabbit normal IgG ab172730 (gray) and 25 µl of Protein A/G Dynabeads. The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach). *http : //www.abcam.com/resources?keywords=X%20ChIP%20protocol

• WB

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Western blot - Anti-Histone H2B (mutated G53D) antibody [EPR26428-35] - BSA and Azide free (AB305053)

This data was developed using ab305052, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Exposure time : 37 seconds

All lanes:

Western blot - Anti-Histone H2B (mutated G53D) antibody [EPR26428-35] (<a href='/en-us/products/primary-antibodies/histone-h2b-mutated-g53d-antibody-epr26428-35-ab305052'>ab305052</a>) at 1/1000 dilution

Lane 1:

HEK-293T cells transfected with a human Histone H2B expression vector containing a his tag, whole cell lysate

Lane 2:

HEK-293T cells transfected with a human Histone H2B(G53D) expression vector containing a his tag, whole cell lysate

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 14 kDa

Observed band size: 14 kDa

false

Exposure time: 37s