Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

Cleaved Histone H3 (Thr22) Antibody #7469

Applications Reactivity Sensitivity MW (kDa) Source
W H X (M) (R) (Mk) (B) (Dg) Recombinant 15 Rabbit

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  X=Xenopus  B=Bovine  Dg=Dog
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Cleaved Histone H3 (Thr22) Antibody detects recombinant or enriched endogenous histone H3 protein only when cleaved in vitro with Cathepsin L at Thr22.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Thr22 of human histone H3 protein. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of acid-extracted histones from NTERA-2 cells (lanes 1 and 2) and recombinant Xenopus histone H3 (lanes 3 and 4), undigested (-) or digested in vitro with Cathepsin L (+), using Cleaved Histone H3 (Thr22) Antibody (upper) or Histone H3 (D1H2) XP® Rabbit mAb #4499 (lower).

Background

Modulation of chromatin structure has a critical role in the control of various DNA directed activities such as transcription, DNA replication, and repair (1). The basic unit of chromatin, the nucleosome, consists of two turns of DNA wrapped around two copies each of four core histone proteins (H2A, H2B, H3, and H4) (2,3). Amino-terminal tails of histones undergo various post-translational modifications such as acetylation, methylation, phosphorylation, and ubiquitination in response to physiological and environmental stimuli. These modifications modulate the accessibility of chromatin to effector proteins as well as act as binding sites for specific histone modification recognizing effector proteins that regulate gene expression (1,4,5). Such alterations in chromatin modifications and architecture that accompany gene expression changes have been observed during embryonic stem cell differentiation (6). One of the ways in which chromatin modifications may be altered in stem cells involves regulated proteolysis of histone H3 by Cathepsin L. Cathepsin L cleaves the histone H3 amino-terminal tail predominantly at Thr22 in differentiating stem cells, leading to removal of histone modification marks which could then influence the expression patterns of developmentally regulated genes (7).

  1. Smith, E. and Shilatifard, A. (2010) Mol Cell 40, 689-701.
  2. Kornberg, R.D. (1974) Science 184, 868-71.
  3. Kornberg, R.D. and Lorch, Y. (1999) Cell 98, 285-94.
  4. Strahl, B.D. and Allis, C.D. (2000) Nature 403, 41-5.
  5. Gardner, K.E. et al. (2011) J Mol Biol 409, 36-46.
  6. Young, R.A. (2011) Cell 144, 940-54.
  7. Duncan, E.M. et al. (2008) Cell 135, 284-94.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

Products