Antibodies, Proteins, Kits and Reagents for Life Science

314 937 ₸ Product size

100 µg 314 937 ₸

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Anti-Histone H3 (phospho S10) antibody [mAbcam 14955] (ab14955)

Key features and details

Mouse monoclonal [mAbcam 14955] to Histone H3 (phospho S10)
Suitable for: IHC-P, ELISA, WB
Reacts with: Human, Drosophila melanogaster, Recombinant fragment
Isotype: IgG1

Product name

Anti-Histone H3 (phospho S10) antibody [mAbcam 14955]
See all Histone H3 primary antibodies
Description

Mouse monoclonal [mAbcam 14955] to Histone H3 (phospho S10)
Host species

Mouse
Specificity

ab14955 recognises phospho S10 on Histone H3. It recognises a phospho S28 peptide by ELISA, but is not blocked by phospho S28 in WB.

Tested Applications & Species

Application	Species
ELISA	Recombinant fragment
IHC-P	Human
WB	Human Drosophila melanogaster

See all applications and species data

Immunogen

Synthetic peptide within Human Histone H3 aa 1-100 (phospho S10) conjugated to keyhole limpet haemocyanin. The exact sequence is proprietary.
(Peptide available as ab11477, ab15644)
Positive control
- WB: Control HeLa Histone Prep. Colcemid treated HeLa Histone prep. HeLa cell lysate treated with calyculin A. IHC-P: Human kidney tissue.
General notes

This antibody clone is manufactured by Abcam.

If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Hybridomas were prepared and the resulting clones were positively screened by ELISA against the immunising tri methyl K9 and phospho S10 dimodified peptide. Clones were also positively screened against both tri methyl K9 and phospho S10 peptides. Clones were negatively screened against the unmodified version of the peptides. This clone binds to the tri methyl K9 and phospho S10 dimodified peptide and to the phospho S10 peptide, but not to the tri methyl K9 peptide or to equivalent unmodified Histone H3 peptide.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.50
Preservative: 0.02% Sodium azide
Constituent: PBS
Concentration information loading...
Purity

IgG fraction
Clonality

Monoclonal
Clone number

mAbcam 14955
Isotype

IgG1
Research areas

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (phospho S10) antibody [mAbcam 14955] (ab14955) Ibañez Rodriguez et al PLoS One. 2016 Nov 18;11(11):e0167063. doi: 10.1371/journal.pone.0167063. eCollection 2016. Fig 4. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

Confocal microscopy of sagittal sections through the rat pineal gland (PG) immunolabeled for Pax6 (green) and the mitotic M-phase marker phospho-histone H3 (PH3, ab14955, red). All PH3+ mitotic cells expressed Pax6; Pax6 levels, however, varied among dividing pinealocyte precursor cells. The number of mitotic precursor cells increased progressively during embryogenesis, and dropped quickly after birth. White arrowheads: Pax6^high mitoses. White arrows: Pax6^low mitoses.

E, embryonic day. PC, posterior commissure. P, postnatal day. SCO, subcommissural organ. 3v, third ventricle.
Western blot - Anti-Histone H3 (phospho S10) antibody [mAbcam 14955] (ab14955)

All lanes : Anti-Histone H3 (phospho S10) antibody [mAbcam 14955] (ab14955) at 1 µg/ml

Lane 1 : Control HeLa Histone Prep 0.5ug
Lane 2 : Colcemid treated HeLa Histone prep 0.5ug
Lane 3 : Control HeLa Histone Prep 0.5ug with Human Histone H3 (tri methyl K9, phospho S10) peptide (ab15644) at 1 µg/ml
Lane 4 : Colcemid treated HeLa Histone prep 0.5ug with Human Histone H3 (tri methyl K9, phospho S10) peptide (ab15644) at 1 µg/ml
Lane 5 : Control HeLa Histone Prep 0.5ug with Human Histone H3 (unmodified) peptide (ab7228) at 1 µg/ml
Lane 6 : Colcemid treated HeLa Histone prep 0.5ug with Human Histone H3 (unmodified) peptide (ab7228) at 1 µg/ml
Lane 7 : Control HeLa Histone Prep 0.5ug with Human Histone H3 (phospho S10) peptide (ab11477) at 1 µg/ml
Lane 8 : Colcemid treated HeLa Histone prep 0.5ug with Human Histone H3 (phospho S10) peptide (ab11477) at 1 µg/ml
Lane 9 : Control HeLa Histone Prep 0.5ug with Human Histone H3 (phospho S28) peptide (ab14793) at 1 µg/ml
Lane 10 : Colcemid treated HeLa Histone prep 0.5ug with Human Histone H3 (phospho S28) peptide (ab14793) at 1 µg/ml

Secondary
All lanes : Rabbit polyclonal to Mouse IgG H&L (HRP) at 1/5000 dilution

Performed under reducing conditions.

Predicted band size: 15 kDa
Observed band size: 17 kDa
why is the actual band size different from the predicted?
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (phospho S10) antibody [mAbcam 14955] (ab14955)

IHC image of ab14955 staining Histone H3 (phospho S10) in human kidney formalin fixed paraffin embedded tissue sections, performed on a Leica Bond.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab14955, 1µg/ml, for 15 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 hematoxylin and mounted with DPX. No primary antibody was used in the negative control (shown on the inset).

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
ELISA - Anti-Histone H3 (phospho S10) antibody [mAbcam 14955] (ab14955)

By ELISA, ab14955 detects:
the singly modified phospho S10 peptide and the dual modified phospho S10 and tri methyl K9 peptide (the orange and grey lines respectively),
less strongly detects the phospho S28 peptide (purple line),
does not detect the equivalent non-modified Histone H3 peptide for S10 or the singly modified tri methyl K9 peptide (the 2 lines at the bottom of the figure).

The antibody recognises phospho S28 by ELISA (although a phospho S28 peptide does not block the antibody in Western blotting).
Western blot - Anti-Histone H3 (phospho S10) antibody [mAbcam 14955] (ab14955)

All lanes : Anti-Histone H3 (phospho S10) antibody [mAbcam 14955] (ab14955) at 1/2000 dilution

Lane 1 : Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate
Lane 2 : HeLa cell lysate treated with calyculin A
Lane 3 : HeLa cell lysate treated with calyculin A and alkaline phosphatase

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP goat anti-rabbit (H+L) at 1/20000 dilution

Predicted band size: 15 kDa

Exposure time: 1 second
Western blot - Anti-Histone H3 (phospho S10) antibody [mAbcam 14955] (ab14955)

All lanes : Anti-Histone H3 (phospho S10) antibody [mAbcam 14955] (ab14955) at 0.5 µg/ml

Lane 1 : Control HeLa Histone prep
Lane 2 : Colecemid treated HeLa Histone prep
Lane 3 : Control HeLa Histone prep with Human Histone H3 (unmodified) peptide (ab7228) at 1 µg/ml
Lane 4 : Colecemid treated HeLa Histone prep with Human Histone H3 (unmodified) peptide (ab7228) at 1 µg/ml
Lane 5 : Control HeLa Histone prep with Human Histone H3 (phospho S28) peptide (ab14793) at 1 µg/ml
Lane 6 : Colecemid treated HeLa Histone prep with Human Histone H3 (phospho S28) peptide (ab14793) at 1 µg/ml
Lane 7 : Control HeLa Histone prep with Human Histone H3 (phospho S10) peptide (ab11477) at 1 µg/ml
Lane 8 : Colecemid treated HeLa Histone prep with Human Histone H3 (phospho S10) peptide (ab11477) at 1 µg/ml

Lysates/proteins at 5 µg per lane.

Secondary
All lanes : Rabbit Anti-Mouse IgG H&L (HRP) (ab6728) at 1/5000 dilution

Predicted band size: 15 kDa
Observed band size: 20 kDa why is the actual band size different from the predicted?
Western blot - Anti-Histone H3 (phospho S10) antibody [mAbcam 14955] (ab14955) Image courtesy of Richelle Sopko, Harvard University, U.S.A

All lanes : Anti-Histone H3 (phospho S10) antibody [mAbcam 14955] (ab14955) at 1/1000 dilution

Lane 1 : Wild type 0-4 hour old fruit fly embryo lysate
Lane 2 : 0-4 hour old fruit fly embryo lysate expressing wee RNAi

Secondary
All lanes : Anti-mouse IgG, peroxidase-linked at 1/10000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 15 kDa

Exposure time: 5 seconds

Blocking: 10% BSA
wee shRNA embryos (lane 2) should display elevated phospho H3Ser10 levels relative to wild type
See Abreview

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