The stoichiometric NADH production can be quantified by spectrophotometric analysis at 440 nm upon reacting with water soluble tetrazolium-1 (WST-1), a soluble tetrazolium dye. The HAT assay does not require radioactivity or chromatography and can be applied for kinetic studies. HATs' counteracting enzymes, histone deacetylases (HDACs), do not interfere with the assay, consequently crude nuclear extract or cell lysate samples can be used.
Additional Materials Required: (Materials Not Included in Kit) Cell lysis buffer (e.g., 1X RIPA buffer) 96-well flat bottom plate.
Product Use: HAT is for research use only. It is not approved for human or animal use, or for application in clinical or in vitro diagnostic procedures.
Shipping and Storage: The product is Dry ice. Upon receipt, components SC8668a, SC8668c, SC8668e, SC8668f and SC8668h should be stored at -20°C, SC8668b, SC8668d and SC8668g should be stored at -80°C. Protect from light. Repeated freeze/thaw cycles should be avoided. Aliquot if necessary. If stored properly, the kit is good for up to 6 months.
Quality Control: Serially diluted HAT positive control samples are measured continuously using this kit for a total of 25 minutes. The detection range of HAT activity is 0.1-2 mUnits/mL.