Store all components (except microtiter plates) at -70°C. Unused HeLa Nuclear Extract should be refrozen quickly, if possible snap freeze in liquid nitrogen.
简单描述
Biocompare Article: Keep an Eye on Apoptosis with Caspase Assays
商品描述
An improved FLUOR DE LYS® HDAC assay with FLUOR DE LYS®-Green, a new substrate offering higher sensitivity and an excitation and emission (485/530nm) that avoids quenching and fluorescent interference from compounds absorbing in the near UV and blue range.The FLUOR DE LYS®-Green HDAC assay is a complete kit for measuring histone deacetylase (HDAC) activity in cell or nuclear extracts, immunoprecipitates or purified enzymes. It comes in a convenient 96-well format, with all reagents necessary for fluorescent HDAC or sirtuin activity measurements and calibration of the assay. The FLUOR DE LYS®-Green HDAC assay is based on the FLUOR DE LYS®-Green substrate and FLUOR DE LYS® developer combination. The FLUOR DE LYS® system (Fluorogenic Histone deAcetylase Lysyl Substrate/Developer) is a highly sensitive and convenient alternative to radiolabeled, acetylated histones or peptide/HPLC methods for the assay of histone deacetylases. The assay procedure has two steps. First, the FLUOR DE LYS®-Green substrate, which comprises an acetylated lysine side chain, is incubated with a sample containing HDAC activity (HeLa nuclear or other extract, purified enzyme, bead-bound immunocomplex, etc.). Deacetylation of the substrate sensitizes the substrate so that, in the second step, treatment with the FLUOR DE LYS® developer produces a fluorophore.
试剂盒组分
Nuclear Extract from HeLa Cells (human cervical cancer cell line) (Prod. No. BML-KI140) (100µl in 0.1M potassium chloride, 20mM HEPES/NaOH, 20% (v/v) glycerol, 0.2mM EDTA, 0.5mM DTT, 0.5mM PMSF) Storage: -70°C. 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 (Prod. No. BML-GR309) (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. NAD+ (Sirtuin Substrate) (Prod. No. BML-KI282) (500µl; 50mM in 50mM TRIS, pH 8.0, 137mM sodium chloride, 2.7mM potassium chloride, 1mM magnesium chloride) Storage: -70°C. Nicotinamide (Prod. No. BML-KI283) (500µl; 50mM in 50mM TRIS, pH 8.0, 137mM sodium chloride, 2.7mM potassium chloride, 1mM magnesium chloride) Storage: -70°C. HDAC Assay Buffer (Prod. No. BML-KI143) (20ml; 50mM TRIS, pH 8.0, 137mM sodium chloride, 2.7mM potassium chloride, 1mM magnesium chloride) Storage: -70°C. 96 Well Plates (Prod. No. BML-KI101) Storage: Room temperature White NBS Microplate (Prod. No. BML-KI571) Storage: Room temperature
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