Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

CHD1 (D8C2) Rabbit mAb #4351

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP ChIP H M R Mk (B) (Dg) (Pg) Endogenous 220 Rabbit IgG

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

Protocols

Specificity / Sensitivity

CHD1 (D8C2) Rabbit mAb recognizes endogenous levels of total CHD1 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human CHD1 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using CHD1 (D8C2) Rabbit mAb.

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 mES cells and either 5 μl of CHD1 (D8C2) Rabbit mAb or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Mouse Oct-4 Promoter Primers #4653, SimpleChIP® Mouse RPL30 Intron 2 Primers #7015, and SimpleChIP® Mouse MYT-1 Promoter Primers #8985. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

Background

Chromodomain-helicase-DNA-binding domain (CHD) proteins have been identified in a variety of organisms (1,2). This family of proteins, which consists of nine members, has been divided into three separate subfamilies: subfamily I (CHD1 and CHD2), subfamily II (CHD3 and CHD4), and subfamily III (CHD5, CHD6, CHD7, CHD8, and CHD9). All of the CHD proteins contain two tandem N-terminal chromodomains, a SWI/SNF-related ATPase domain, and a C-terminal DNA binding domain (1,2). The chromodomains facilitate binding to methylated lysine residues of histone proteins and confer interactions with specific regions of chromatin. The SWI/SNF-related ATPase domain utilizes the energy from ATP hydrolysis to modify chromatin structure. CHD1 is a euchromatic protein that associates with the promoters of active genes, and is required for the maintenance of open chromatin and pluripotency in embryonic stem cells (3-6). The two chromodomains of CHD1 facilitate its recruitment to active genes by binding to methyl-lysine 4 of histone H3, a mark associated with transcriptional activation (4-6). Yeast CHD1 is a component of the SAGA and SLIK histone acetyltransferase complexes, and is believed to link histone methylation with histone acetylation during transcriptional activation (6). The CHD2 protein is not well characterized; however, mouse knockout studies suggest important functions in development and tumor suppression. Homozygous CHD2 knockout mice exhibit delayed growth and perinatal lethality (7). Heterozygous knockout mice show increased mortality and gross organ abnormalities, in addition to increased extramedullary hematopoiesis and susceptibility to lymphomas (7,8). CHD2 mutant cells are defective in hematopoietic stem cell differentiation and exhibit aberrant DNA damage responses (8).

  1. Hall, J.A. and Georgel, P.T. (2007) Biochem Cell Biol 85, 463-76.
  2. Marfella, C.G. and Imbalzano, A.N. (2007) Mutat Res 618, 30-40.
  3. Gaspar-Maia, A. et al. (2009) Nature 460, 863-8.
  4. Sims, R.J. et al. (2005) J Biol Chem 280, 41789-92.
  5. Flanagan, J.F. et al. (2005) Nature 438, 1181-5.
  6. Pray-Grant, M.G. et al. (2005) Nature 433, 434-8.
  7. Marfella, C.G. et al. (2006) J Cell Physiol 209, 162-71.
  8. Nagarajan, P. et al. (2009) Oncogene 28, 1053-62.

Application References

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