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Applications: Useful for the study of bromodomain binding assays, screening bromodomain inhibitors and selectivity profiling.
Contraindications: DMSO above 0.5%. Only limited amounts of DMSO can be included, as it has been shown to disrupt BRD-ligand interaction. Avoid green and blue dyes that absorb light in the Alphascreen® signal emission range (520-620 nm), such as Trypan Blue. Avoid the use of the potent singlet oxygen quenchers such as sodium azide (NaN3) or metal ions (Fe2+, Fe3+, Cu2+, Zn2+ and Ni2+). The presence of >1% RPMI 1640 culture medium leads to a signal reduction due to the presence of excess biotin and iron in this medium. MEM, which lacks these components, does not affect Alphascreen® assays.
Description: The CREBBP Inhibitor Screening Assay Kit is designed to measure the inhibition of CREBBP binding to its substrate. The key to the CREBBP Inhibitor Screening Assay Kit is a specific binding of the CREBBP bromodomain to the acetylated histone substrate. With this kit, only three simple steps on a microtiter plate are required. First, a sample containing CREBBP and an inhibitor of choice is incubated with the biotinylated substrate for thirty minutes. Next, acceptor beads are added, then donor beads, followed by reading the Alpha-counts.
Storage Stability: At least 6 months from date of receipt, when stored as directed. Kit components require different storage conditions. Be sure to store each component at the proper temperature upon arrival.
Supplied As: Kit comes in a convenient AlphaLISA® format, with biotinylated histone peptide substrate, assay buffer, detection buffer and purified GST-tagged CREBBP bromodomain to perform a total of 384 enzyme reactions.
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: Filippakopoulos, P., et al., Cell 2012; 149:214.