All lanes : Anti-Histone H2A (phospho T120) antibody [EPR17492] (ab177391) at 1/1000 dilution

Lane 1 : Core histones purified by acid extraction precipitation from untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) cells
Lane 2 : Core histones purified by acid extraction precipitation from HeLa (Human epithelial cell line from cervix adenocarcinoma) cells treated with colcemid

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 14 kDa
Observed band size: 14 kDa


Exposure time: 1 second

Blocking/Dilution buffer: 2% BSA/TBST.

This target is specifically expressed at M phase according to the literature [PMID: 19965387].

ab177391 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 human colon tissue labeling Histone H2A (phospho T120) with ab177391 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Sporadically nuclear staining on human colon [PMID: 19965387].

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

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

Immunohistochemical analysis of paraffin-embedded human spleen tissue labeling Histone H2A (phospho T120) with ab177391 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Sporadically nuclear staining on germinal center of human spleen [PMID: 19965387].

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

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

Immunohistochemical analysis of paraffin-embedded human gastric tumor tissue labeling Histone H2A (phospho T120) with ab177391 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Sporadically nuclear staining on tumor cells of human gastric tumor [PMID: 19965387].

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

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

Direct ELISA analysis using ab177391 (0 to 1000 ng/ml) in peptide ELISA with H2A T120 phospho peptide, H2A unmodified peptide, H2A.XT120 phospho peptide and H2A.X unmodified peptide as antigens (at 1000 ng/ml concentration). An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/2500 dilution was used as secondary antibody.