Description:The BRD2 (BD2) Inhibitor Screening Assay Kit is designed to measure the inhibition of BRD2 bromodomain 2 (BD2) from binding to its substrate. The key to the BRD2 (BD2) Inhibitor Screening Assay Kit is the highly specific binding of the BRD2 bromodomain 2 to the acetylated histone substrate. With this kit, only three simple steps on a microtiter plate are required. First, a sample containing BRD2 bromodomain 2 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. Synonym(s): BRD2, RNF3, RING3, Bromodomain Supplied As: Kit comes in a convenient AlphaLISA^® format, with biotinylated histone peptide substrate, assay buffer, detection buffer and purified, GST-tagged BRD2 BD2 to perform a total of 384 enzyme reactions. 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.

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Instructions for use: See assay kit data sheet for detailed protocol. Storage / Stability:

At least 12 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.

Application(s): Useful for the study of bromodomain binding assays, screening bromodomain inhibitors and selectivity profiling. Reference(s): McBride, A.A., et al., Trends Microbiol. 2004; 12(12):527. Notes:

MATERIALS OR INSTRUMENTS REQUIRED BUT NOT SUPPLIED: Glutathione AlphaLISA^® Acceptor Beads, 5 mg/ml (PerkinElmer #AL109C) AlphaScreen^® Streptavidin-conjugated donor beads, 5 mg/ml (PerkinElmer #6760002) Optiplate-384 (PerkinElmer #6007290) AlphaScreen^® microplate reader Adjustable micropipettor and sterile tips

Warning(s): Avoid freeze/thaw cycles. Scientific Category: Bromodomain Product Type: Assay Kit Data shown is lot-specific. Contact us for specific information on other lots.