A semi-automated method for 5'-ectonucleotidase measurement in lymphocytes.
We describe a simple, non-radioactive, semi-automated method for measuring lymphocyte ecto-5'-nucleotidase activity in isolated cell homogenates. The method which uses inosine 5'-monophosphate (IMP) as a substrate was optimized and requires a total of 4 x 10(6) lymphocytes. The reference range obtained (38.7-180.0 nmol/h per mg protein; means = 106.6) agrees with other isotopic methods which use 14C-IMP. The simplicity, sensitivity (lower limit 10, upper limit 400 nmol/h per mL) and precision (at 114 nmol/h per mL, interday 4.2% s.d., n = 20; intraday 5.8%, n = 10) makes this method suitable for routine clinical laboratory use.
Chalmers
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AH
Department of Pathology, Mater Misericordiae Public Hospital, South Brisbane, Australia.
Hare
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C
D015720:5'-Nucleotidase / Q000097:blood*; D000293:Adolescent; D000328:Adult; D000368:Aged; D001331:Automation; D002648:Child; D006801:Humans; D007291:Inosine Monophosphate / Q000378:metabolism; D008214:Lymphocytes / Q000201:enzymology*; D008875:Middle Aged; D012016:Reference Values; D015203:Reproducibility of Results; D013379:Substrate Specificity
D007291:Inosine Monophosphate; D015720:5'-Nucleotidase