Histone H3T11ph antibody (pAb)

RRID: AB_2615078
Catalog No: 39151 Format: 200 µl ¥88,000 Buy
Catalog No: 39152 Format: 10 µl ¥24,000 Buy

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

Applications

ChIP Validated ChIP-Seq Validated Western Blot Validated Dot Blot Validated

Application Notes

Applications Validated by Active Motif:
ChIP: 5 µl per ChIP
ChIP-Seq: 5 µl each
WB*: 1:500 - 1:2,000 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.

Published Applications

The following applications have been published using this antibody. Unless noted above, Active Motif may not have validated the antibody for use in these applications:

WB

View publications that use Active Motif products & services here. Enter the product number(s) to see publications & applications that use this antibody.

Immunogen

This Histone H3 phospho Thr11 antibody was raised against a peptide including phospho-threonine 11 of histone H3.

Buffer

Rabbit serum containing 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic. For your convenience, an IgG version (Catalog No. 61197) of this antibody that was purified by Protein A Chromatography is also available.

View our Guide to Histone Modifications and Biological Function.

 

Histone H3 phospho Thr11 pAb tested by ChIP-Seq.
ChIP was performed using the ChIP-IT® High Sensitivity Kit (Cat. No. 53040) with chromatin from the human H9 embryonic stem cell line (4.5 million cells) and 5 ul of antibody. ChIP DNA was sequenced on the Illumina HiSeq and 22 million sequence tags were mapped to identify H3T11ph occupancy. The image on the left shows H3T11ph binding at transcriptional start sites across a 120,000 bp region on chromosome 1. The image on the right is zoomed in to show H3T11ph binding at a single promoter

.Histone H3 phospho Thr11 pAb tested by Western blot.
Western blot probed with Histone H3 phospho Thr11 pAb (1:2,000 dilution).
    Lane 1: 200 ng recombinant histone H3.
    Lane 2: 5 µg acid extract of HeLa cells.
    Lane 3: 5 µg acid extract of HeLa cells treated with colcemid.

Histone H3 phospho Thr11 pAb tested by dot blot analysis.
Dot blot analysis was used to confirm the specificity of Histone H3 phospho Thr11 pAb for phospho-Thr11 of histone H3. Decreasing amounts of peptides corresponding to regions around major sites of histone H3 threonine phosphorylation were spotted onto PVDF and probed with the antibody at a dilution of 1:10,000.
    Lane 1: Peptide phosphorylated at threonine 3.
    Lane 2: Unmodified threonine 3 peptide.
    Lane 3: Peptide phosphorylated at threonine 11.
    Lane 4: Unmodified threonine 11 peptide.
    Lane 5: Peptide phosphorylated at threonine 45.
    Lane 6: Unmodified threonine 45 peptide.

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.

Phosphorylation of several residues of histone H3, such as Ser10 (Histone H3 phospho Ser10), Ser28 (Histone H3 phospho Ser28) and Thr11, is tightly correlated with chromosome condensation during both mitosis and meiosis. Localization of histone H3 phospho Thr11 is different in mammals and in plants. In plant cells, phosphorylation of Thr11 of histone H3 is distributed along the entire length of condensed chromosome. In mammals, this modification is restricted to the centromeric region of the chromosome. Thr11 of histone H3 is phosphorylated by Dlk/ZIP kinases during mitosis.

Positive Control

Active Motif's HeLa acid extract (PTX treated) (Catalog No. 36201) can be used as a positive control for Histone H3 phospho Thr11 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 H3T11ph 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