DESCRIPTION

• 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
• Shiptemp: -80°C (dry ice)
• Synonyms: BRD3, RING3L, Bromodomain
• Warnings: Avoid freeze/thaw cycles.
• Category: Bromodomain/Assay Kit
• References: Muller S, Filippakopoulos P, Knapp S. Expert Rev Mol Med. 2011 Sep 13;13:e29.
• Description: The BRD3 (BD2) Inhibitor Screening Assay Kit is designed to measure the inhibition of BRD3 bromodomain 2 (BD2) from binding to its substrate. The key to the BRD3 (BD2) Inhibitor Screening Assay Kit is the highly specific binding of the BRD3 bromodomain 2 to the acetylated histone substrate. With this kit, only three simple steps on a microtiter plate are required. First, a sample containing BRD3 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.
• Methodology: AlphaLISA®
• Supplied As: Kit comes in a convenient AlphaLISA® format, with biotinylated histone peptide substrate, assay buffer, detection buffer and purified, GST-tagged BRD3 BD2 to perform a total of 384 enzyme reactions.
• Unspsc Code: 41121811
• Unspsc Name: Laboratory microchemistry kits
• Applications: Useful for the study of bromodomain binding assays, screening bromodomain inhibitors and selectivity profiling.
• Product Type: Assay Kit
• Biosafety Level: Not applicable (BSL-1)
• 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.
• 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.
• Scientific Category: Bromodomain
• Instructions for Use: See assay kit datasheet for detailed protocol.