Histone H3S28ph antibody (mAb)

RRID: AB_2732838
Clone: 5F9A9
Catalog No: 61695 Format: 100 µg ¥5,500 Buy
Catalog No: 61696 Format: 10 µg ¥1,380 Buy

Request a quote for a bulk order Request Quote


Product Review Submit a Review


Antibody Type:
Monoclonal
Isotype:
IgG2b
Purification:
Protein A Chromatography
Host:
Mouse
Molecular Weight:
17 kDa
Reactivity:
Human, Wide Range Predicted

Applications

Western Blot Validated Immunofluorescence Validated Immunocytochemistry Validated

Application Notes

Applications Validated by Active Motif:
ICC/IF: 2 - 10 µg/ml dilution
WB*: 0.5 - 2 µg/ml 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 antibody was raised against a peptide corresponding to phospho-serine 28 of human Histone H3.

Buffer

Purified IgG in PBS with 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic.

View our Guide to Histone Modifications and Biological Function.

 

Histone H3S28ph antibody (mAb) (Clone 5F9A9) tested by Immunofluorescence.
Left: Formaldehyde fixed mitotic DM4 cell stained with Histone H3S28ph antibody (mAb).
Right: Hoechst stain.

Histone H3S28ph antibody (mAb) (Clone 5F9A9) tested by Western blot.
Detection of Histone H3S28ph antibody by Western blot. The analysis was performed using 20 µg of untreated (lane 1) or colcemid treated (lane 2) HeLa nuclear extract with Histone H3S28ph antibody at a 1 µg/ml dilution.

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. In contrast to Ser10 phosphorylation, Ser28 phosphorylation has never been observed in interphase.

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