Anti-Histone H2B (mutated G53D) antibody [EPR26428-35]

4 product images

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  ChIP - Anti-Histone H2B (mutated G53D) antibody [EPR26428-35] (ab305052)

  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 Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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

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  Western blot - Anti-Histone H2B (mutated G53D) antibody [EPR26428-35] (ab305052)

  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] (ab305052) 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) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Predicted band size: 14 kDa

  Observed band size: 14 kDa

  Exposure time: 37s

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  Immunocytochemistry - Anti-Histone H2B (mutated G53D) antibody [EPR26428-35] (ab305052)

  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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

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  Flow Cytometry (Intracellular) - Anti-Histone H2B (mutated G53D) antibody [EPR26428-35] (ab305052)

  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, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution was used as the secondary antibody.