A quantitative method for measurement of lysosomal acid phosphatase latency in cultured rat heart cells with 210Pb.
A method is described for measuring the latency of lysosomal acid phosphatase in cultured rat heart endotheloid cells. 210Pb was added to a medium used to demonstrate acid phosphatase activity by the Gomori lead method, and the amount of lead deposited was measured with a liquid scintillation counter. Deposition rates were measured after enzyme activation pretreatments with acetate buffer (pH 5.0) at various osmolalities, and after formaldehyde fixation. Formaldehyde, alloxan, or fluoride in the Gomori medium were evaluated for their differential effects on lysosomal and non-lysosomal acid phosphatase. The method was found to provide a sensitive, rapid and quantitative evaluation of acid phosphatase latency and should be useful for studying the integrity of lysosomes within cells.
D000085:Acetates / Q000494:pharmacology; D000135:Acid Phosphatase / Q000378:metabolism*; D000496:Alloxan / Q000494:pharmacology; D000818:Animals; D046508:Culture Techniques; D004727:Endothelium; D004789:Enzyme Activation / Q000187:drug effects; D005459:Fluorides / Q000494:pharmacology; D005557:Formaldehyde / Q000494:pharmacology; D006651:Histocytochemistry / Q000379:methods*; D007854:Lead; D008247:Lysosomes / Q000201:enzymology*; D009206:Myocardium; D009994:Osmolar Concentration; D011868:Radioisotopes; D051381:Rats
D000085:Acetates; D011868:Radioisotopes; D005557:Formaldehyde; D007854:Lead; D000496:Alloxan; D000135:Acid Phosphatase; D005459:Fluorides