All lanes : Anti-Histone H3 (mono methyl K14) antibody [EPR17708-235] (ab202416) at 1/1000 dilution

Lane 1 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate
Lane 2 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate

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


Exposure time: 1 minute

This data was developed using ab202416, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer: 5% NFDM/TBST.

This data was developed using ab202416, the same antibody clone in a different buffer formulation.Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Histone H3 (mono methyl K14) with ab202416 at 1/2000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).Confocal image showing nuclear staining on HeLa cell line.The nuclear counter stain is DAPI (blue).Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).The negative controls are as follows:
-ve control 1: ab202416 at 1/2000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

This data was developed using ab202416, the same antibody clone in a different buffer formulation.Flow cytometry analysis of HeLa cells labelling Histone H3 (mono methyl K14) (red) with purified ab202416 at dilution of 1/12 000. The secondary antibody used was Alexa Fluor^® 488 goat-anti-rabbit IgG (1/2000). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Isotype control antibody used was Rabbit monoclonal IgG (black). The blue line shows cells without incubation with primary antibody and secondary antibody.

This data was developed using ab202416, the same antibody clone in a different buffer formulation.ab202416 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.