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A semi‐automated method for 5′‐ectonucleotidase measurement in lymphocytes
Author(s) -
Chalmers A. H.,
Hare Cavelyn
Publication year - 1990
Publication title -
immunology and cell biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.999
H-Index - 104
eISSN - 1440-1711
pISSN - 0818-9641
DOI - 10.1038/icb.1990.11
Subject(s) - chemistry , computer science , microbiology and biotechnology , biology
Summary We describe a simple, non‐radioactive, semi‐automated method for measuring lymphocyteccto‐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 4×10 6 lymphocytes. The reference range obtained (38·7–180·0 nmol/h per mg protein; = 106·6) agrees with other isotopic methods which use 14 C‐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.