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Erythrocyte contamination of leukocyte populations following density‐gradient centrifugation results in artificially high levels of human leukocyte HMG‐CoA reductase activity
Author(s) -
Harwood H. James,
Bridge Donna M.,
Stacpoole Peter W.
Publication year - 1988
Publication title -
lipids
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.601
H-Index - 120
eISSN - 1558-9307
pISSN - 0024-4201
DOI - 10.1007/bf02535283
Subject(s) - reductase , lysis , centrifugation , differential centrifugation , biochemistry , hmg coa reductase , biology , enzyme , chemistry , chromatography , microbiology and biotechnology
When measuring human leukocyte HMG‐CoA reductase activity, special care must be taken to prevent erythrocyte contamination of the leukocyte layer during isopycnic centrifugation. Contamination during leukocyte isolation and subsequent erythrocyte lysis during NH 4 Cl treatment results in increased leukocyte microsomal HMG‐CoA reductase activity. Increased enzyme activity is not due to enzyme dephosphorylation, thiol‐disulfide reduction or increased enzyme protein concentration. Leukocyte populations containing granulocytes appear to be most sensitive. Prevention of erythrocyte contamination during isopycnic centrifugation should aid in accurate measurement of human leukocyte HMG‐CoA reductase activity.