Recombinant KAT2B / PCAF protein

Contents

A representative Technical Data Sheet (TDS) is provided here. Please refer to the lot-specific TDS you will receive with your order for the lot-specific buffer contents and protein concentration.

Background

p300/CBP-associated factor (PCAF), also known as K (lysine) acetyltransferase 2B (KAT2B), is a transcriptional coactivator associated with p53 that contains acetyltransferase and E3 ubiquitin ligase domains as well as a bromodomain for interaction with other proteins. The bromodomain functions as a ‘reader’ of epigenetic histone marks and regulates chromatin structure and gene expression by linking associated proteins to the recognized acetylated nucleosomal targets. PCAF competes with E1A for binding to p300/CBP, which associate with various sequence-specific factors involved in cell growth and/or differentiation, including c-Jun and the adenoviral oncoprotein E1A. PCAF exerts histone acetyltransferase activity in association with core histones (H3 and H4) and with nucleosome core particles, and functions to promote transcriptional activation. PCAF has also been shown to inhibit cell cycle progression and to counteract the mitogenic activity of the adenoviral oncoprotein E1A. In case of HIV-1 infection, PCAF is recruited by the viral protein TAT to regulate TAT’s transactivating activity and may help induce chromatin remodeling of proviral genes.

Application Notes

This protein is suitable for use in binding assays, inhibitor screening, and selectivity profiling.

Assay Conditions: 0.5 µg Histone H4 (Cat. No. 31493) was incubated with 0 µg (-), 0.5 µg (+) PCAF in a 20 µl reaction system containing 50 mM Tris-HCl pH 8.0, 0.1 mM EDTA, 50 ng/µl BSA, 1 mM TCEP and 20 µM Acetyl-CoA for 2 hours at room temperature. Half of each reaction was run on a 13% SDS-PAGE gel, and products were detected by Western blot. H4ac (pan-acetyl) antibody (Cat. No. 39243) was used to recognize acetylated histone H4.

Protein Details

Recombinant human PCAF was expressed in a baculovirus expression system as the full length protein (accession number NP_003875.3) with an N-terminal FLAG-Tag. The molecular weight of the protein is 94.3 kDa.

References

The following applications have been published using this protein. Unless noted above, Active Motif may not have validated this product for use in these applications:

Acetylation Assay

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Storage

Recombinant proteins in solution are temperature sensitive and must be stored at -80°C to prevent degradation. Avoid repeated freeze/thaw cycles and keep on ice when not in storage.

Guarantee

This product is for research use only and is not for use in diagnostic procedures. This product is guaranteed for 6 months from date of arrival.