05-806
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
NACRES:
NA.41
eCl@ss:
32160702
Clone:
3H10, monoclonal
Species reactivity:
human
Application:
FACS, ICC, IF, WB, inhibition assay, multiplexing
Citations:
92
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.
17 kDa
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:
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:
Biochem/physiol Actions
Broad species cross-reactivity is expected.
Histone H3 phosphorylated at Ser10.
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.
Preparation Note
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.
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.
UV-treated 293 cell extracts, UV-treated HeLa cell extracts, breast cancer tissue, HEPG2 cell extracts.
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).
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).
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
WGK 1
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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