p300 Tools and Services for Research and Drug Discovery
Active Motif offers a workflow of research tools and approaches to characterize the complex functions of p300/CBP.
p300 and CBP (CREB Binding Protein) are transcriptional co-activators that regulate gene expression through acetylation of histones and chromatin remodeling. p300 interacts with over 50 intracellular proteins affecting a range of signaling pathways, while mutations in p300 can lead to dysregulated cell growth and division, and are implicated in acute myeloid leukemia (AML) and multiple solid tumor types.
- Recombinant Proteins, Enzymes, and Antibodies
- Assays, Kits, and Small Molecule Inhibitors
- ChIP-Seq Services and RNA-Seq Services
Featured Publication: Raisner, R., et. al. 2018. “Enhancer Activity Requires CBP/P300 Bromodomain-Dependent Histone H3K27 Acetylation.” Cell Reports 24: 1722-1729.
(Active Motif products used in this study: Recombinant p300 protein, Recombinant CREBBP (CBP), Histone PTM antibodies, and RIME services).
Recombinant p300 proteins – Histone Acetyltransferase Activity with Histone H4 Substrate
Activity of Recombinant p300 proteins was assessed using the Active Motif HAT Assay Kit. Equimolar amounts of p300, Catalytic Domain (Cat. No. 81093) and p300, Full Length (81158) proteins were incubated with H4 peptide for 30 minutes to 6 hours, and developed as per the kit protocol. Graph shows averages ± SEM of arbitrary fluorescence units (AFU).
Recombinant p300 proteins – Histone Acetyltransferase Activity with Histone H3 Substrate
Activity of p300, Catalytic Domain (Cat. No. 81093), p300, Full Length (Cat. No. 81158), and p300, Bromodomain (Cat. No. 31372) was assessed using the Active Motif HAT Assay Kit. Equimolar amounts of p300 proteins were incubated with H3 peptide for 3 hours, and developed as per the kit protocol. Anacardic acid, an inhibitor of HAT activity, was used at 15μM / well. Graph shows averages ± SEM of arbitrary fluorescence units (AFU).
Histone Acetyltransferase Activity of Recombinant p300 Proteins by Western Blot
Recombinant Histone H4 (Cat. No. 31493) was incubated with 0 μg, 0.25 μg or 0.5 μg of p300 proteins in a 30 μl reaction system including 50 mM Tris pH 8.0, 1 mM TCEP, 0.1 mM EDTA, 50 ng/μl BSA and 20 μM acetyl-CoA for 30 minutes at 30°C. For each reaction, 7.5 μl of reaction product was run on a 12.5% SDS-PAGE gel and detected by Western Blot using Histone H4 pan-acetyl antibody (Cat. No. 39243).
p300 antibody (mAb) tested by ChIP-Seq
ChIP was performed using the ChIP-IT® High Sensitivity Kit (Cat. No. 53040) with chromatin from 4.5 million LNCaP cells and 5 µl of p300 antibody (Cat. No. 61401). ChIP DNA was sequenced on the Illumina HiSeq and 15 million sequence tags were mapped to identify p300 binding sites. p300 along with H3K4me1 are markers of active enhancer elements and are therefore expected to co-localize. A sampling of the p300 ChIP-Seq data shows the expected co-localization of p300 and H3K4me1.
|Name||Expressed In||Format||Cat. No.|
|Recombinant p300 protein||Baculovirus||10 µg||81158|
|Recombinant p300 protein, catalytic domain||Baculovirus||20 µg||81093|
|Recombinant p300 (1041-1161) protein||E. coli||100 µg||31372|
|Recombinant CREBBP (1075-1873) protein||Baculovirus||20 µg||31590|
|Recombinant CREBBP (1081-1197) protein||E. coli||100 µg||31373|
|Recombinant KAT2A (GCN5) protein||Baculovirus||20 µg||31591|
|Recombinant KAT2B / PCAF protein||Baculovirus||20 µg||81142|
|Recombinant KAT2B / PCAF (715-829) protein||E. coli||100 µg||31370|
|Recombinant p53 (TP53) protein||Baculovirus||20 µg||81091|
|p300 antibody (mAb)||IgG1||Human||ChIP, ChIP-Seq, ICC, IF, IP, WB||100 µg||61401|
|AbFlex® Histone H3K27ac antibody (rAb)||IgG2a||Human||ChIP-Seq, ELISA, WB||100 µg||91193|
|Histone H3K27ac antibody (pAb)||IgG||Budding Yeast, Human, Other (Wide Range)||ChIP, ChIP-Seq, ICC, IF, IP, WB||100 µg||39133|
|Histone H3K27ac antibody (mAb)||IgG1||Human, Other (Wide Range)||ChIP, ChIP-Seq, DB, ICC, IF, WB||100 µg||39685|
|RNA pol II antibody (mAb)||IgG2a||Human||ChIP, ChIP-Seq, ICC, IF, WB||100 µg||61667|
|HAT Assay Kit (Fluorescent)||1 x 96 rxns||56100|