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.
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.
Materials Required But Not Supplied
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
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.
Instructions for Use
See assay kit data sheet for detailed protocol.
Applications
Useful for the study of bromodomain binding assays, screening bromodomain inhibitors and selectivity profiling.
Shipping Temperature
dry ice
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
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.
Warnings
Avoid freeze/thaw cycles.
Disclaimers and Limitations
BPS Bioscience assay kits are validated using the listed components according to our specific protocol. Any deviations to kit components or protocols, such as using different proteins, cell/tissue lysates, or buffers (including using the same component from other commercial sources) will void any warranties on the performance of the assay kit and is not recommended.
End-users should immediately report any issues to [email protected] upon receipt of the kit, such as missing components, damaged vials, no dry ice, etc.