Cell Signaling Technology

Product Pathways - Chromatin Regulation

PCAF (C14G9) Rabbit mAb #3378

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP ChIP H M R Mk (B) (Hr) Endogenous 93 Rabbit igG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  ChIP=Chromatin IP
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  B=Bovine  Hr=Horse
Species cross-reactivity is determined by Western blot.

Protocols

Specificity / Sensitivity

PCAF (C14G9) Rabbit mAb detects endogenous levels of total PCAF protein. The antibody does not cross-react with the related GCN5L2 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the human PCAF protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using PCAF (C14G9) Rabbit mAb.

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 293 cells treated with Forskolin #3828 (30 µM) and either 20 μl of PCAF (C14G9) Rabbit mAb #3378 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 primers specific for the ALS2 and NR4A3 genes, and the heterochromatic α Satellite repeat element. 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

p300/CBP-associated factor (PCAF), also known as lysine acetyl-transferase 2B (KAT2B), is a transcriptional adaptor protein and histone acetyl-transferase (HAT) that functions as the catalytic subunit of the PCAF transcriptional co-activator complex (1). PCAF is 73% homologous to GCN5L2, another HAT protein found in similar complexes (1,2). Like GCN5L2, PCAF acetylates histone H3 on Lys14 and histone H4 on Lys8, both of which contribute to gene activation by modulating chromatin structure and recruiting additional co-activator proteins that contain acetyl-lysine binding bromo-domains (3). PCAF also acetylates non-histone proteins including transcriptional activators (p53, E2F1, MyoD), general transcription factors (TFIIEβ and TFIIF) and architectural DNA binding proteins (HMGA1 and HMG17) (4-10). Acetylation of these proteins regulates their nuclear localization, protein stability, DNA binding, and co-activator association.

  1. Nagy, Z. and Tora, L. (2007) Oncogene 26, 5341-57.
  2. Yang, X.J. et al. (1996) Nature 382, 319-24.
  3. Schiltz, R.L. et al. (1999) J Biol Chem 274, 1189-92.
  4. Bannister, A.J. and Miska, E.A. (2000) Cell Mol Life Sci 57, 1184-92.
  5. Liu, L. et al. (1999) Mol Cell Biol 19, 1202-9.
  6. Martínez-Balbás, M.A. et al. (2000) EMBO J 19, 662-71.
  7. Sartorelli, V. et al. (1999) Mol Cell 4, 725-34.
  8. Imhof, A. et al. (1997) Curr Biol 7, 689-92.
  9. Munshi, N. et al. (1998) Mol Cell 2, 457-67.
  10. Herrera, J.E. et al. (1999) Mol Cell Biol 19, 3466-73.

Application References

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Companion Products

Rabbit monclonal antibody is produced under license (granting certain rights including those under U. S. Patent No. 5,675,063) from Epitomics, Inc.

This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

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