Rabbit Anti-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.
Applications
Suitable for use in Western Blot and Immunoprecipitation. Other applications not tested.
Recommended Dilutions
Western 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. Immunoprecipitation: 5-10ug immunoprecipitates PCAF from RIPA lysate with ~4x10e7 human acute T cell leukemia Jurkat cells. Optimal dilutions to be determined by the researcher.
Positive Antigen Control
Unstimulated 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.
Storage and Stability
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Immunogen
Synthetic peptide [EKFFFSKIKEAGLIDK] (with cysteine added to N-terminal) conjugated to KLH, corresponding to the C-terminal amino acids 817-832 of human PCAF. Species sequence homology: mouse and chicken
Form
Supplied as a liquid in PBS, pH 7.4, 15mM sodium azide.
Purity
Purified by affinity chromatography.
Specificity
Recognizes human PCAF. Species Crossreactivity: canine
