Histone H3S10phS28ph antibody (pAb)

RRID: AB_2793167
Catalog No: 39147 Format: 200 µl ¥5,830 Add to Cart
Catalog No: 39148 Format: 10 µl ¥1,430 Add to Cart

Request a quote for a bulk order Request Quote

Product Review Submit a Review

Antibody Type:
Polyclonal
Isotype:
Serum
Purification:
None
Host:
Rabbit
Molecular Weight:
17 kDa
Reactivity:
Human, Wide Range Predicted

Applications

ChIP Validated Western Blot Validated Immunofluorescence Validated Immunocytochemistry Validated

Application Notes

Applications Validated by Active Motif:
ChIP: 10 µl per ChIP
ICC/IF: 1:500 - 1:2,000 dilution
WB*: 1:1,000 - 1:10,000 dilution

*Note: many chromatin-bound proteins are not soluble in a low salt nuclear extract and fractionate to the pellet. Therefore, we recommend a High Salt / Sonication Protocol when preparing nuclear extracts for Western blot.

Immunogen

This Histone H3 phospho Ser10,28 antibody was raised against a peptide including phosphorylated serine 28 of human Histone H3.

Buffer

Rabbit serum containing 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic.

View our Guide to Histone Modifications and Biological Function.

 
Histone H3S10phS28ph antibody (pAb) tested by ChIP.

Histone H3 phospho Ser10,28 antibody tested by ChIP analysis.
Chromatin IP performed using the ChIP-IT® Express Kit (Catalog No. 53008) and HeLa Chromatin (1.5 x 106 cell equivalents per ChIP) using 10 µl of Histone H3 phospho Ser10, 28 antibody or the equivalent amount of rabbit IgG as a negative control. Real time, quantitative PCR (RT-qPCR) was performed on DNA purified from each of the ChIP reactions using a primer pair specific for the GAPDH gene promoter. Data are presented as Fold Enrichment of the ChIP antibody signal versus the negative control IgG using the ddCT method.

Histone H3S10phS28ph antibody (pAb) tested by immunofluorescence.

Histone H3 phospho Ser10,28 antibody tested by immunofluorescence.
HeLa cells stained with Histone H3 phospho Ser10,28 antibody at a 1:1,000 dilution.
    Top panel: Histone H3 phospho Ser10,28 antibody.
    Middle: DAPI.
    Bottom: Merge of both images.

Histone H3S10phS28ph antibody (pAb) tested by Western blot.

Histone H3 phospho Ser10,28 antibody tested by Western blot.
HeLa acid extract probed with Catalog No. 39147 (1:5,000 dilution).
    Lane 1: No treatment.
    Lane 2: Cells treated with colcemid.

Background

Histone H3 is one of the core components of the nucleosome. The nucleosome is the smallest subunit of chromatin and consists of 147 base pairs of DNA wrapped around an octamer of core histone proteins (two each of Histone H2A, Histone H2B, Histone H3 and Histone H4). Chromatin is subject to a variety of chemical modifications, including post-translational modifications of the histone proteins and the methylation of cytosine residues in the DNA. Reported histone modifications include acetylation, methylation, phosphorylation, ubiquitylation, glycosylation, ADP-ribosylation, carbonylation and SUMOylation; these modifications play a major role in regulating gene expression.

Ser10 phosphorylation and Ser28 phosphorylation in the tail of H3 have very similar kinetics. Both phosphorylations occur early in mitosis when chromosomes begin to condense, and during premature chromosome condensation induced in S-phase cells. These phosphorylated serines are excellent mitotic markers.

Positive Control

Active Motif's HeLa nuclear extract (Catalog No. 36010) can be used as a positive control for Histone H3 phospho Ser10,28 antibody.

Chromatin IP Assays

This antibody has been validated for use in ChIP and/or ChIP-Seq, and can be used with Active Motif's ChIP-IT® High Sensitivity Kit or our magnetic bead-based ChIP-IT® Express Kits. For an overview of all of our ChIP products, please visit our Chromatin IP Page to learn about ChIP products designed for use with Histone H3S10phS28ph antibody (pAb).

Storage

Some products may be shipped at room temperature. This will not affect their stability or performance. Avoid repeated freeze/thaw cycles by aliquoting items into single-use fractions for storage at -20°C for up to 2 years. Keep all reagents on ice when not in storage.

Guarantee

This product is guaranteed for 12 months from date of receipt.

This product is for research use only and is not for use in diagnostic procedures.

Application Key

  • ChIP = Chromatin Immunoprecipitation;
  • ChIP = ChIP Sequencing;
  • CUT&RUN = Cleavage Under Targets and Release Using Nuclease;
  • CUT&Tag = Cleavage Under Targets and Tagmentation;
  • DB = Dot Blot;
  • ELISA = Enzyme-linked Immunosorbent Assay;
  • EMSA = Electrophoretic Mobility Shift Assay
  • FC = Flow Cytometry;
  • ICC = Immunocytochemistry;
  • IF = Immunofluorescence;
  • IHC = Immunohistochemistry;
  • IP = Immunoprecipitation;
  • MeDIP = Methyl-DNA Immunoprecipitation;
  • NEU = Neutralizing;
  • WB = Western Blotting