Anti-Histone H3 (tri methyl K9, phospho S10) antibody

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (tri methyl K9, phospho S10) antibody (AB5819)

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.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K9, phospho S10) antibody (AB5819)

ab5819 staining Histone H3 (tri methyl K9, phospho S10) in mitotic HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab5819 at 0.5μg/ml and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 594), at 2μg/ml (shown in red). The secondary antibody (shown in green) was ab150081, Alexa Fluor® 488 Goat anti-Rabbit IgG (H+L) used at a 1/1000 dilution for 1h at room temperature. Nuclear DNA was labelled with DAPI (shown in blue).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• ICC/IF

AbReview4483****

Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K9, phospho S10) antibody (AB5819)

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.

• ICC/IF

Collaborator

Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K9, phospho S10) antibody (AB5819)

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.

This image is courtesy of Darin McDonald, Hendzel Laboratory

• WB

Lab

Western blot - Anti-Histone H3 (tri methyl K9, phospho S10) antibody (AB5819)

Blocking buffer : 2% BSA

Gel type : MES

All lanes:

Western blot - Anti-Histone H3 (tri methyl K9, phospho S10) antibody (ab5819) at 1 µg/mL

Lane 1:

Calf Thymus Histone at 0.5 µg

Lane 2:

HeLa Nuclear – Triton Prep at 10 µg

Lane 3:

NIH3T3 Nuclear – Triton Prep at 10 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution

Predicted band size: 15 kDa

Observed band size: 17 kDa

false

Exposure time: 2min

• WB

Unknown

Western blot - Anti-Histone H3 (tri methyl K9, phospho S10) antibody (AB5819)

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).

All lanes:

Western blot - Anti-Histone H3 (tri methyl K9, phospho S10) antibody (ab5819) at 1/500 dilution

Lanes 1, 5, 7 and 9:

HeLa histones at 0.5 µg

Lanes 2, 6, 8 and 10:

Colcemid-treated HeLa histones at 0.5 µg

Lane 3:

HeLa histones at 0.5 µg with Human Histone H3 (tri methyl K9, phospho S10) peptide (<a href='/en-us/products/unavailable/human-histone-h3-tri-methyl-k9-phospho-s10-peptide-ab15644'>ab15644</a>)

Lane 4:

Colcemid-treated HeLa histones at 0.5 µg with Human Histone H3 (tri methyl K9, phospho S10) peptide (<a href='/en-us/products/unavailable/human-histone-h3-tri-methyl-k9-phospho-s10-peptide-ab15644'>ab15644</a>)

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab6721'>ab6721</a>) at 1/5000 dilution

Predicted band size: 15 kDa

Observed band size: 22 kDa,60 kDa

false