Isopropanol Dehydrogenase Activity Colorimetric Assay Kit provides a convenient tool for sensitive detection of the ADH1- NADP in a variety of samples. In this assay, Isopropanol gets converted to Acetone and NADPH in the presence of NADP+ . This results in the development of colour in the presence of the developer. This colour is proportional to the ADH1-NADP levels in the sample and can be measured at λ = 450 nm. This assay detects ADH1-NADP activity as low as 0.001 mU in samples.
NADP-dependent Isopropanol Dehydrogenase (ADH1-NADP, EC:18.104.22.168) belongs to the superfamily of alcohol dehydrogenases (Alcohol DHs, ADHs, EC 22.214.171.124) with a preference for medium chain secondary alcohols, such as 2-butanol and isopropanol. Isopropanol dehydrogenase facilitates the conversion between secondary alcohols to aldehydes and ketones with the reduction of NADP+ to NADPH.
Measurement of Isopropanol Dehydrogenase (ADH1-NADP) activity in various cells.Measurement of NADP-dependent alcohol dehydrogenases" activity with broad substrate specificity in various tissues/cells.
Mammalian samples. Bacterial, yeast or protozoan samples: Entamoeba histolytica, Clostridium beijerinckii, Mycoplasma pneumonia, etc.
Store the kit at -20°C, protected from light. Briefly centrifuge vials prior to opening. Read the entire protocol before performing the assay.ADH1-NADP Assay Buffer: Warm to room temperature before use. Store at either 4ºC or -20ºC.ADH1-NADP Developer: Reconstitute the Developer with 0.9 ml of ddH2O. Pipette up and down several times to completely dissolve the pellet (Do not vortex). Aliquot and store at –20°C. Keep on ice while in use. Use within two months.ADH1-NADP Positive Control: Dilute the Positive Control with 100 µl Assay Buffer and mix thoroughly. Aliquot and store directly at -20°C. Keep on ice while in use. Use within two months.NADPH Standard: Reconstitute NADPH Standard with 200 µl pure DMSO to generate 1 mM NADPH Standard solution. Aliquot and store at –20°C. Keep on ice while in use. Use within two months.
ADH1-NADP Assay Buffer: 25 mlADH1-NADP Substrate: 1 mlADH1-NADP Developer: 1 vialADH1-NADP Positive Control: 10 µlNADPH Standard: 1 vial
Absorbance (450 nm)
Compatible Sample Types:
• Bacterial, yeast or protozoan samples: Entamoeba histolytica, Clostridium beijerinckii, Mycoplasma pneumonia, etc.• Animal tissues: such as liver, pancreas, small intestine, etc. • Animal cells.
Features & Benefits:
• Simple, colorimetric protocol.• Detects ADH1-NADP activity as low as 0.001 mU in samples.• Compatible with multiple sample types.