Determination of acid phosphatase activity in macrophages / 中国免疫学杂志

ABSTRACT

A simple and accurate method is described for the quantitative determination of acid phosphatase in macrophage, spleen and thymus homogenates, utilizing sodium ?-naphthyl acid phosphate as substrate, and the monocoupler Fast Red B Salt as the color developer to measure ?-naphthol released from the substrate by enzymatic hydrolysis reaction.The color product is a deep blue azo dye,stable for at least 1h in alkaline condition. The method is sensitive enough for assay of enzyme activity of macrophages at concentration low as 1?10~6 cell/ml. The acid phosphatase activity of macrophages from mice treated with thioglycollate elevated significantly. Some factors affecting assay are discussed. Above all, the color developer solution should be made freshly and the peritoneal macrophages collected from mice should be broken in an ultrasonic breaker before assay.