PCAF (p300/CBP-associated factor, CREBBP-associated factor, CAF, P/CAF)

A direct link exists between hyperacetylation of chromatin and transcriptional activation. Experiments have established that PCAF possesses intrinsic histone acetylase activity. PCAF primarily acetylates lysine 14 of H3. It acetylates lysine 8 of H4 less efficiently. PCAF also acetylates p53 in response to DNA damage. The distinct patterns of acetylation by PCAF may contribute to transcriptional regulation of important genes.\n\nApplications: \nSuitable for use in Western Blot and Immunoprecipitation. Other applications not tested.\n\nRecommended Dilution:\nWestern Blot: 1:500; Determined using whole extracts of human epidermoid carcinoma A431, canine MDCK kidney cells or recombinant active human PCAF, and a chemiluminescent detection reagent. Predicted molecular weight: ~88kD. In some extract preparations, additional band(s) may appear. Staining of PCAF band(s) by immunoblotting is inhibited by the PCAF immunizing peptide.\nImmunoprecipitation: 5ug immunoprecipitates PCAF from RIPA lysate with ~4x10e7 human acute T cell leukemia Jurkat cells.\nOptimal dilutions to be determined by the researcher.\n\nPositive Antigen Control: \nUnstimulated A431 cell lysate. Add 2.5ul of 2-mercaptoethanol/100ul of lysate and boil for 5 minutes to reduce the preparation. Load 20ug of reduced lysate per lane for minigels.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage, store at -20 degrees C. Aliquots are stable for at least 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

P3114-05A

Size

100ul

Applications

IP, WB

Hosts

Rabbit

Reactivities

Hum, Can

Form

Supplied as a liquid in PBS, pH 7.4, 1% BSA, 15mM sodium azide, before the addition of glycerol to 40%.

P Type

Pab

Purity

Purified by immunoaffinity chromatography.

Isotype

IgG

References

1. Roth, S.Y., et al., Annu. Rev. Biochem. 70: 81-120 (2001) PubMed: 11395403. 2. Marmorstein, R., Cell Mol. Life Sci. 58: 693-703 (2001) PubMed: 11437231. 3. Sterner, D.E. & Berger, S.L., Microbiol. Mol. Biol. Rev. 64: 435-459 (2000) PubMed: 10839822. 4. Kouzarides, T., EMBO J 19: 1176-1179 (2000) PubMed: 10716917.

Additional Info

Recognizes human and canine PCAF. Species Crossreactivity: Predicted to react with mouse and chicken due to sequence homology.