Anti-Histone H4 (acetyl K12) antibody [EPR17906]

7 product images

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  Western blot - Anti-Histone H4 (acetyl K12) antibody [EPR17906] (ab177793)

  Blocking/Dilution buffer: 5% NFDM/TBST.

  All lanes: Western blot - Anti-Histone H4 (acetyl K12) antibody [EPR17906] (ab177793) at 1/10000 dilution

  Lane 1: HeLa (Human epithelial cells from cervix adenocarcinoma) treated with 500 ng/ml Trichostatin A for 4 hours whole cell lysates at 10 µg

  Lane 2: HeLa (Human epithelial cells from cervix adenocarcinoma) untreated whole cell lysates at 10 µg

  Secondary

  All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

  Predicted band size: 11 kDa

  Observed band size: 11 kDa

  Exposure time: 15s

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  Immunocytochemistry/ Immunofluorescence - Anti-Histone H4 (acetyl K12) antibody [EPR17906] (ab177793)

  Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells, treated with Trichostatin A (500 ng/ml) for 4 hours or untreated, labeling Histone H4 (acetyl K12) with ab177793 at 1/2000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing nuclear staining on HeLa cell line is observed. Acetylation level increased after treatment with Trichostatin A (500 ng/ml) for 4 hours. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
  The negative controls are as follows;
  1. ab177793 at 1/2000 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
  2. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

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  Flow Cytometry (Intracellular) - Anti-Histone H4 (acetyl K12) antibody [EPR17906] (ab177793)

  Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) treated (Red)/untreated (Green) with 500ng/ml Trichostatin A for 4 hours with purified ab177793 at 1/80 dilution. The secondary antibody was Goat anti rabbit IgG (Alexa Fluor^® 488) at 1/2000 dilution. A Rabbit monoclonal IgG (Black) was used as the isotype control and cells without incubation with primary antibody and secondary antibody (Blue) were used as unlabeled control.
  

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  Peptide Array - Anti-Histone H4 (acetyl K12) antibody [EPR17906] (ab177793)

  ab177793 was tested in Peptide array against 501 different modified and unmodified histone peptides; each peptide is printed on the array at six concentrations (each in triplicate).
  Circle area represents affinity between the antibody and a peptide: all antigen-containing peptides are displayed as red circles, all other peptides as blue circles. The affinity is calculated as area under curve when antibody binding values are plotted against the corresponding peptide concentration. Each circle area is normalized to the peptide with the strongest affinity.
  The complete dataset, including full list of all peptides and information on the position of each peptide in the diagram, is available under the Support & downloads section.

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  Western blot - Anti-Histone H4 (acetyl K12) antibody [EPR17906] (ab177793)

  Blocking/Dilution buffer: 5% NFDM/TBST.

  All lanes: Western blot - Anti-Histone H4 (acetyl K12) antibody [EPR17906] (ab177793) at 1/10000 dilution

  Lane 1: NIH/3T3 (Mouse embyro fibroblast cells) treated with 500 ng/ml Trichostatin A for 4 hours whole cell lysates at 10 µg

  Lane 2: NIH/3T3 (Mouse embyro fibroblast cells) untreated whole cell lysates at 10 µg

  Secondary

  All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

  Predicted band size: 11 kDa

  Exposure time: 15s

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H4 (acetyl K12) antibody [EPR17906] (ab177793)

  Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Histone H4 (acetyl K12) with ab177793 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500. Nucleus staining on Human colon tissue is observed. Counter stained with Hematoxylin.
  

  Negative control: Using PBS instead of primary antibody.

  Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H4 (acetyl K12) antibody [EPR17906] (ab177793)

  Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling Histone H4 (acetyl K12) with ab177793 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500. Nucleus staining on Rat colon tissue is observed. Counter stained with Hematoxylin.
  

  Negative control: Using PBS instead of primary antibody.

  Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.