Histone H3T3ph antibody (pAb)

RRID: AB_2793170
Catalog No: 39153 Format: 200 µl ¥86,000 Buy
Catalog No: 39154 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

Western Blot Validated Immunofluorescence Validated Dot Blot Validated Immunocytochemistry Validated

Application Notes

Applications Validated by Active Motif:
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:

ICC
IF

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 Thr3 antibody was raised against a peptide including phospho-threonine 3 of 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 H3 phospho Thr3 pAb tested by Western blot.
Western blot probed with Histone H3 phospho Thr3 pAb (1:500 dilution).
    Lane 1: 20 µg acid extract of HeLa cells.
    Lane 2: 20 µg acid extract of HeLa cells treated with colcemid.

Histone H3 phospho Thr3 pAb tested by dot blot analysis.
Dot blot analysis was used to confirm the specificity of Histone H3 phospho Thr3 pAb for phospho-Thr3 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 1:2,000.
    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.

In vivo, phosphorylation of histone H3 at threonine 3 begins at early prophase in the vicinity of the nuclear envelope, spreads to the pericentromeric chromatin during prometaphase and is fully reversed by late anaphase, 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 Thr3 antibody.

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