Filter paper disk techniques for assay of nucleotidase.
A DE filter disk technique for assaying the activity of nucleotidase is described. This method is based on the observation that nucleotides bind to the filters at 5 mM Tris-HCl (pH 7.8) while nucleosides do not. As parameter for the nucleotidase activity the decrease of bound nucleotides is determined. In parallel experiments the amount of the product (nucleoside) formed can be measured by DEAE Sephadex column chromatography. The filter disk technique can be applied for the determination of vmax and Km of a nucleotidase by using different ribonucleosidase monophosphate substrates.
D000249:Adenosine Monophosphate / Q000378:metabolism; D000818:Animals; D002417:Cattle; D007700:Kinetics; D008722:Methods; D009705:Nucleosides / Q000302:isolation & purification; D009708:Nucleotidases / Q000032:analysis* / Q000378:metabolism; D009711:Nucleotides / Q000302:isolation & purification; D010209:Paper; D013379:Substrate Specificity; D013950:Thymus Gland / Q000201:enzymology
D009705:Nucleosides; D009711:Nucleotides; D000249:Adenosine Monophosphate; D009708:Nucleotidases