FLUOR DE LYS®-Green HDAC2 fluorometric drug discovery assay kit

Store all components except the microtiter plate and instruction booklet at -70°C for the highest stability. HDAC2 (Prod. No. BML-KI575) must be handled with particular care in order to retain maximum enzymatic activity. Defrost it quickly in a RT water bath or by rubbing between fingers, then immediately store on an ice bath. The remaining unused enzyme should be refrozen quickly, by placing at -70°C. If possible, snap freeze in liquid nitrogen or a dry ice/ethanol bath. To minimize the number of freeze/thaw cycles, aliquot the enzyme into separate tubes and store at -70°C. The FLUOR DE LYS®-Green Substrate (Prod. No. BML-KI572), when diluted in Assay Buffer, may precipitate after freezing and thawing. It is best, therefore, to dilute only the amount needed to perform the assays of that day.

The original FLUOR DE LYS ® HDAC Fluorescent Activity Assay revolutionized HDAC and sirtuin screening by providing a highly sensitive, mix-and-read alternative to radiolabeled, acetylated histones or peptide/HPLC methods for the assay of lysine deacetylases. The FLUOR DE LYS ® -Green assay has those same advantages, but also, due to its higher wavelength excitation and emission, avoids interference by quenching or fluorescence from compounds absorbing and/or emitting in the near UV and blue. FLUOR DE LYS ® -Green is an especially sensitive substrate for HDAC2.

HDAC2 (Histone Deacetylase 2) (human, recombinant) (Prod. No. BML-KI575) (67μl; 30ng/μl in 50mM TRIS, pH 8.0, 138mM NaCl, 10% glycerol, 1mg/ml BSA) Storage: -70°C. Avoid freeze/thaw cycles! FLUOR DE LYS ® -Green substrate (Prod. No. BML-KI572) (50μl; 50mM in DMSO) Storage: -70°C FLUOR DE LYS ® Developer concentrate (20x) (Prod. No. BML-KI105) (300μl; 20x Stock solution; Dilute in Assay Buffer before use.) Storage: -70°C Trichostatin A (HDAC Inhibitor) (Prod. No. BML-GR309-9090) (100μl; 0.2mM in DMSO) Storage: -70°C FLUOR DE LYS ® -Green Standard (Prod. No. BML-KI605) (30μl 1mM in DMSO) Storage: -70°C. Note: BML-KI605 is an improved replacement of BML-KI573. HDAC Assay Buffer II (Prod. No. BML-KI422) (20ml; 50mM TRIS/Cl, pH 8.0, 137mM NaCl, 2.7mM KCl, 1mM MgCl 2 , 1mg/ml BSA) Storage: -70°C 1/2 volume black NBS microplate (Prod. No. BML-KI570) Storage: Room temperature 1/2 volume white NBS microplate (Prod. No. BML-KI571) Storage: Room temperature