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Key Documents

05-806

Sigma-Aldrich

Anti-phospho-Histone H3 (Ser10) Antibody, clone 3H10

clone 3H10, Upstate®, from mouse

Synonym(s):

H3 histone, family 3A, H3S10P, Histone H3 (phospho S10), H3 histone, family 3A, H3 histone, family 3B, H3 histone, family 3B (H3.3B)

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About This Item

UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.41

biological source

mouse

Quality Level

antibody form

purified immunoglobulin

antibody product type

primary antibodies

clone

3H10, monoclonal

species reactivity

human

manufacturer/tradename

Upstate®

technique(s)

flow cytometry: suitable
immunocytochemistry: suitable
immunofluorescence: suitable
inhibition assay: suitable (peptide)
multiplexing: suitable
western blot: suitable

isotype

IgG1κ

NCBI accession no.

UniProt accession no.

shipped in

wet ice

target post-translational modification

phosphorylation (pSer10)

Gene Information

human ... HIST1H3F(8968)

General description

Histone H3 is one of the five main histone proteins involved in the structure of chromatin in eukaryotic cells. Featuring a main globular domain and a long N-terminal tail, H3 is involved with the structure of the nucleosomes of the ′beads on a string′ structure. The N-terminal tail of histone H3 protrudes from the globular nucleosome core and can undergo several different types of epigenetic modifications that influence cellular processes. These modifications include the covalent attachment of methyl or acetyl groups to lysine and arginine amino acids and the phosphorylation of serine or threonine.

Specificity

Broad species cross-reactivity is expected.
Histone H3 phosphorylated at Ser10.

Application

Anti-phospho-Histone H3 (Ser10) Antibody, clone 3H10 is a mouse monoclonal antibody for detection of Histone H3 phosphorylated at serine 10. This purified mAb, also known as H3S10p, is published in peer reviewed journals and has been validated in FC, ICC, IF, PIA, WB, Mplex.
Flow Cytometry:
This antibody has been reported by an independent laboratory to detect phosphorylated histone H3 using flow cytometry.

Beadlyte Histone-Peptide Specificity Assay:
1:1,000 to 1:81,000 dilutions of a previous lot were incubated with histone H3 peptides containing various modifications conjugated to Luminex microspheres. (Figure B). Only the peptide containing phospho-serine 10 was detected.


Immunocytochemistry:
0.2 μg/mL of a previous lot showed positive chromosome immunostaining for mitotic A431 and HeLa cells fixed with 95% ethanol and 5% acetic acid and permeabilized with 0.1% Triton X-100.

Peptide Inhibition Analysis:
Detection of histone H3 in immunoblots of colcemid treated HeLa acid extracts by anti-phospho-Histone H3 (Ser10) was diminished by 10 μM of histone H3 peptide containing phospho-serine 10, but not by peptides containing phospho-serine 28 or an unmodified histone H3 sequence (Figure C).

Immunofluorescence:

Quality

Routinely evaluated by western blot acid extracted proteins from mitotic HeLa cells treated with colcemid (Catalog # 17-306).

Western Blot Analysis:
0.05-0.5 μg/mL of this lot detected phosphorylated histone H3 in acid extracted proteins from mitotic HeLa cells treated with colcemid (Catalog # 17-306), but not unmodified recombinant Histone H3 (Catalog # 14-494) (Figure A).

Target description

17 kDa

Physical form

Format: Purified
Purified mouse IgG1k in buffer containing 0.1 M Tris-glycine, pH 7.4, 0.15 M NaCl and 0.05% sodium azide.

Storage and Stability

Stable for 1 year at 2 to 8°C from date of receipt.

Handling Recommendations:
Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. Note: Variability in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.

Analysis Note

Control
UV-treated 293 cell extracts, UV-treated HeLa cell extracts, breast cancer tissue, HEPG2 cell extracts.

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Legal Information

UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

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Storage Class

12 - Non Combustible Liquids

wgk_germany

WGK 1

flash_point_f

Not applicable

flash_point_c

Not applicable


Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Maomao Zhang et al.
PloS one, 10(3), e0119285-e0119285 (2015-03-06)
The spindle assembly checkpoint (SAC) maintains the fidelity of chromosome segregation during mitosis. Nonpathogenic cells lacking the SAC are typically only found in cleavage stage metazoan embryos, which do not acquire functional checkpoints until the mid-blastula transition (MBT). It is
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May, M; Schelle, I; Brakebusch, C; Rottner, K; Genth, H
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Florian Ulrich et al.
Developmental biology, 357(1), 134-151 (2011-07-13)
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Christopher Runyan et al.
Development (Cambridge, England), 133(24), 4861-4869 (2006-11-17)
During germ-cell migration in the mouse, the dynamics of embryo growth cause many germ cells to be left outside the range of chemoattractive signals from the gonad. At E10.5, movie analysis has shown that germ cells remaining in the midline

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