Rabbit polyclonal to Histone H3 (phospho S10, tri methyl K9) - ab5819, on histone preparations from HeLa cells. Primary antibody used at 1/500.

Lanes 1, 3, 5, 7, 9 - control HeLa histones (0.5ug/lane)
Lanes 2, 4, 6, 8, 10 - Colcemid-treated HeLa histones (0.5ug/lane)

All blocking peptides are 1ug/lane

Lanes 1 and 2: ab5819  
Lanes 3 and 4: ab5819 + Histone H3 (Tri Me K9, phospho S10) peptide - ab15644
Lanes 5 and 6: ab5819 + Histone H3 peptide (unmodified) - ab7228
Lanes 7 and 8: ab5819 + Histone H3  (phospho S10) peptide - ab11477
Lanes 9 and 10: ab5819 + Histone H3 (tri methyl K9) peptide - ab1773 

Secondary antibody - Goat polyclonal to rabbit IgG (HRP) - ab6721 1/5000.

ab5819 is specific for Histone H3 phosphorylated at residue S10 and tri methylated at residue K9. The activity of the antibody is blocked by the addition of the immunizing peptide, ab15644 (lanes 3 and 4).

SKN cells stained with ab5819 (green) at a dilution of 1/2000. The cells were fixed in paraformaldehyde for 10 minutes prior to incubation with ab5819. The DNA is stained with DAPI (blue). 100x magnification.

ab5819 at a 1/500 dilution staining mitotic human HeLa cells by immunocytochemistry. The antibody was incubated with formaldehyde fixed cells for 1 1/2 hours and bound antibody was then detected using a Texas red conjugated goat anti-rabbit IgG.

This image is courtesy of an Anonymous Abreview submitted on 6 April 2006.

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IHC image of Histone H3 (tri methyl K9, phospho S10) staining in human breast carcinoma FFPE section, performed on a Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab5819, 5µg/ml, for 8 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.