1% for the planet logo

Phospho-Histone H3 (Ser28) Antibody #9713

Filter:
  • WB
  • IF
  • F
Western blot analysis of lysates from CHO and HeLa cells either untreated or synchronized in metaphase by treatment with 100 ng/ml nocodazole for 4 h, followed by isolation of metaphase cells by mitotic shake-off. Blots were probed with Phospho-Histone H3 (Ser28) Antibody #9713 (upper) or Histone H3 Antibody #9715 (lower).

To Purchase # 9713

Supporting Data

REACTIVITY H M Hm Dm
SENSITIVITY Endogenous
MW (kDa) 17
SOURCE Rabbit
Application Key:
  • WB-Western Blotting 
  • IF-Immunofluorescence 
  • F-Flow Cytometry 
Species Cross-Reactivity Key:
  • H-Human 
  • M-Mouse 
  • Hm-Hamster 
  • Dm-D. melanogaster 
  • Related Products

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunofluorescence (Frozen) 1:400 - 1:1600
Immunofluorescence (Immunocytochemistry) 1:400 - 1:1600
Flow Cytometry (Fixed/Permeabilized) 1:200 - 1:800

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.

Protocol

Specificity / Sensitivity

Phospho-Histone H3 (Ser28) Antibody detects endogenous levels of histone H3 only when phosphorylated at Ser28. This antibody does not cross-react with other phosphorylated histones, including phospho-histone H3 (Ser10). Non-specific cytoplasmic staining may be observed by immunofluorescence in fixed-frozen tissues.

Species Reactivity:

Human, Mouse, Hamster, D. melanogaster

The antigen sequence used to produce this antibody shares 100% sequence homology with the species listed here, but reactivity has not been tested or confirmed to work by CST. Use of this product with these species is not covered under our Product Performance Guarantee.

Species predicted to react based on 100% sequence homology:

Rat, Chicken, Xenopus, Zebrafish, Bovine

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the amino terminus of histone H3 phosphorylated on Ser28. Antibodies are purified by protein A and peptide affinity chromatography.

Background

Modulation of chromatin structure plays an important role in the regulation of transcription in eukaryotes. The nucleosome, made up of DNA wound around eight core histone proteins (two each of H2A, H2B, H3, and H4), is the primary building block of chromatin (1). The amino-terminal tails of core histones undergo various posttranslational modifications, including acetylation, phosphorylation, methylation, and ubiquitination (2-5). These modifications occur in response to various stimuli and have a direct effect on the accessibility of chromatin to transcription factors and, therefore, gene expression (6). In most species, histone H2B is primarily acetylated at Lys5, 12, 15, and 20 (4,7). Histone H3 is primarily acetylated at Lys9, 14, 18, 23, 27, and 56. Acetylation of H3 at Lys9 appears to have a dominant role in histone deposition and chromatin assembly in some organisms (2,3). Phosphorylation at Ser10, Ser28, and Thr11 of histone H3 is tightly correlated with chromosome condensation during both mitosis and meiosis (8-10). Phosphorylation at Thr3 of histone H3 is highly conserved among many species and is catalyzed by the kinase haspin. Immunostaining with phospho-specific antibodies in mammalian cells reveals mitotic phosphorylation at Thr3 of H3 in prophase and its dephosphorylation during anaphase (11).

Pathways

Explore pathways related to this product.


For Research Use Only. Not for Use in Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
All other trademarks are the property of their respective owners. Visit our Trademark Information page.