2x HDAC Assay Developer is a reagent designed to be used with BPS Bioscience HDAC (histone deacetylase) or sirtuin fluorogenic assay kits containing Fluorogenic HDAC/SIRT Substrate 1 (BPS Bioscience #50032), Fluorogenic HDAC Class 2a Substrate (BPS Bioscience #50040), Fluorogenic HDAC Class 2a Substrate 2 (Green) (BPS Bioscience #50042), Fluorogenic HDAC Substrate 2 (Green) (BPS Bioscience #50038) and Fluorogenic HDAC Substrate 3 (BPS Bioscience #50037). Use of an appropriate fluorogenic substrate, containing acetylated lysine side chain, and incubation with a sample containing HDAC activity (purified enzyme) results in deacetylation of the substrate. This deacetylation of the substrate allows the generation of a fluorescent signal when incubated with HDAC Assay Developer. Trichostatin A is included in the formulation to stop Class 1 and 2 HDACs reactions and allow measurement of HDAC activity at the time of addition.
1. K. Ito et al. Proc. Natl. Acad. Sci. USA 2002 99 8921. 2. B. Heltweg and M. Jung Anal. Biochem. 2002 302 175. 3. S. Milutinovic et al. J. Biol. Chem. 2002 277 20974
Storage/Stability
Upon receipt, store at -80°C. Concentrate is stable for at least 3 freeze-thaw cycles, but care should be taken that both freezing and thawing steps are done quickly (use liquid nitrogen or dry ice/EtOH for freezing). Frozen storage of smaller size aliquots is recommended. When stored at -80°C, the product is stable for 15 months from date of receipt. Avoid repeated freeze/thaw cycles.
Applications
Study of HDAC or sirtuin kinetics, regulation and inhibitor sensitivity in drug discovery and high through put screening (HTS) biochemical and cell based HDAC assays. It is suitable for use with BPS Bioscience HDAC (histone deacetylase) or sirtuin fluorogenic assay kits containing Fluorogenic HDAC/SIRT Substrate 1 (BPS Bioscience #50032), Fluorogenic HDAC Class 2a Substrate (BPS Bioscience #50040), Fluorogenic HDAC Class 2a Substrate 2 (Green) (BPS Bioscience #50042), Fluorogenic HDAC Substrate 2 (Green) (BPS Bioscience #50038) or Fluorogenic HDAC Substrate 3 (BPS Bioscience #50037). Use in other assays may require optimization.