Information

A FLUOR DE LYS^® fluorescent assay system. The HDAC6 Fluorescent Activity Assay/Drug Discovery Kit is a complete assay system designed to measure the lysyl deacetylase activity of the recombinant human HDAC6 included in the kit. The kit is ideal for chemical library screening for candidate inhibitors or activators or kinetic assay of the enzyme under varying conditions. The FLUOR DE LYS^® HDAC6 assay is based on the FLUOR DE LYS^® Substrate and FLUOR DE LYS^®Developer combination. The assay procedure has two steps. First, the FLUOR DE LYS^® Substrate, which comprises an acetylated lysine side chain, is incubated with HDAC6. Deacetylation of the substrate sensitizes the substrate so that, in the second step, treatment with the FLUOR DE LYS^®Developer produces a fluorophore. 

Alternative Name:	Histone deacetylase 6 fluorescent assay kit
Applications:	Fluorescent detection, HTS Activity assay, Cell-based assays
Use/Stability:	Store all components except the microplate and instruction booklet at -70°C for the highest stability. The HDAC6 Enzyme (Prod. No. BML-SE508), 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.
Shipping:	Shipped on Dry Ice
Long Term Storage:	-80°C
Contents:	HDAC6 (Histone Deacetylase 6) (human, recombinant) (Prod. No. BML-SE508) (50 µg; 10mM TRIS, pH 7.5, 100mM sodium chloride, 3mM magnesium chloride, 10% glycerol) Storage: -70°C, avoid freeze/thaw cycles! FLUOR DE LYS® SIRT1, Deacetylase Substrate (Prod. No. BML-KI177) (100µl; 5mM solution in 50mM TRIS/Cl, pH 8.0, 137mM sodium chloride, 2.7mM potassium chloride, 1mM magnesium chloride) Storage: -70°C FLUOR DE LYS® Developer II Concentrate (5x) (Prod. No. BML-KI176) (5 x 250 µl; 5x Stock Solution; Dilute in Assay Buffer before use Storage: -70°C Trichostatin A (HDAC Inhibitor) (Prod. No. BML-GR-309-9090) (100 µl; 0.2mM in DMSO) Storage: -70°C FLUOR DE LYS® Deacetylated Standard (Prod. No. BML-KI142) (30 µl; 10mM in DMSO) Storage: -70°C HDAC Assay Buffer II (Prod. No. BML-KI422) (20ml; 50mM TRIS/Cl, pH 8.0, 137mM sodium chloride, 2.7mM potassium chloride, 1mM magnesium chloride, 1mg/ml bovine serum albumin) Storage: -70°C HDAC Assay Buffer (Prod. No. BML-KI143) (20ml; 50mM TRIS/Cl, pH 8.0, 137mM sodium chloride, 2.7mM potassium chloride, 1mM magnesium chloride) Storage: -70°C 1/2 volume microplate (Prod. No. BML-KI101) Storage: Room temperature 1/2 volume white NBS microplate (Prod. No. BML-KI571) Storage: Room temperature
Scientific Background:	HDAC6 is a class II HDAC, a group defined by its member’s homology to the yeast HDAC. HDAC6 falls into the class IIb subclass, along with HDAC10, but is unique among human HDACs in that it contains two full deacetylase domains. Primarily located to cytoplasm, HDAC6 is a tubulin deacetylase. Other HDAC6 substrates include Hsp90, cortactin and peroxiredoxins. Through these deacetylase activities and its ubiquitin and other binding activities, HDAC6 plays key regulatory roles in cell motility, protein folding, the ubiquitin-proteasome pathway, the aggresome pathway and autophagy, suggesting a central function in the coordination of cellular stress responses. HDAC6 is required for efficient oncogenic tumorigenesis. The mechanisms underlying this requirement may include HDAC6-mediated promotion of invasive cell motility, resistance to anoikis, removal of toxic misfolded proteins, inhibition of Hsp90 stabilization of metastasis suppressors and promotion of angiogenesis. HDAC6 null-mice, while outwardly normal, have increased resistance to chemical carcinogenesis. It is thus reasonable to suppose that HDAC6 inhibition may be a key factor in the anti-cancer effects of pan-HDAC inhibitors and that ive HDAC6 inhibitors may have the potential for greater effectiveness and reduced side-effects. Manual  / MSDS