All lanes : Anti-Histone H3 (acetyl K36) antibody [EPR16992] (ab177179) at 1/1000 dilution

Lane 1 : HeLa treated with Sodium butyrate extract lysates
Lane 2 : Untreated Hela extract lysates

Lysates/proteins at 10 µg per lane.

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

Predicted band size: 15 kDa

Blocking/Dilution buffer: 5% NFDM/TBST.

All lanes : Anti-Histone H3 (acetyl K36) antibody [EPR16992] (ab177179) at 1/1000 dilution

Lane 1 : NIH/3T3 treated with Trichostatin A whole cell lysates
Lane 2 : Untreated NIH/3T3 whole cell lysates

Lysates/proteins at 10 µg per lane.

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

Predicted band size: 15 kDa
Observed band size: 15 kDa

Blocking/Dilution buffer: 5% NFDM/TBST.

Immunohistochemical analysis of paraffin-embedded Human skin tissue labeling Histone H3 (acetyl K36) with ab177179 at 1/100 dilution, followed by prediluted Goat Anti-Rabbit IgG H&L (HRP). Nucleus staining on Human skin tissue is observed. Counter stained with Hematoxylin.
Negative control: Using PBS instead of primary ab, secondary ab is prediluted Goat Anti-Rabbit IgG H&L (HRP).

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

ab177179 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, can be downloaded here.

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling Histone H3 (acetyl K36) with ab177179 at 1/100 dilution, followed by prediluted Goat Anti-Rabbit IgG H&L (HRP). Nucleus staining on glandular epithelium of mouse colon tissue is observed. Counter stained with Hematoxylin.
Negative control: Using PBS instead of primary ab, secondary ab is prediluted Goat Anti-Rabbit IgG H&L (HRP).

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

Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling Histone H3 (acetyl K36) with ab177179 at 1/100 dilution, followed by prediluted Goat Anti-Rabbit IgG H&L (HRP). Nucleus staining on glandular epithelium of rat colon tissue is observed. Counter stained with Hematoxylin.

Negative control: Using PBS instead of primary ab, secondary ab is prediluted Goat Anti-Rabbit IgG H&L (HRP).

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

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa cells labeling Histone H3 (acetyl K36) with ab177179 at 1/100 dilution, followed by Goat anti-rabbit Alexa Fluor® 488 (IgG) (ab150077) secondary antibody at 1/400 dilution (green). Nuclear staining on HeLa cell line is observed. The expression increased after treatment with TSA (50 μg/ml) for 4 hours. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and ab150120 (goat anti-mouse AlexaFluor®594 secondary) at 1/500 dilution (red).
The negative controls are as follows;
1. ab177179 at 1/100 dilution followed by ab150120 (goat anti-mouse AlexaFluor®594 secondary) at 1/500 dilution.
2. ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (goat anti-rabbit Alexa Fluor®488 (IgG H&L) at 1/400 dilution.