A microsystem to evaluate the synthesis of [3H]leucine labeled proteins by macrophages.
A method is described for evaluating protein synthesis by adherent M phi by measuring the incorporation of [3H]leucine into TCA precipitable material. By using guanidine-HCl it was possible to remove completely the radiolabeled proteins from the adherent cells that were cultured in microwells, and retain TCA precipitable material. This procedure enabled us to harvest the TCA precipitable proteins with a semiautomatic cell harvester. The guanidine-HCl treatment did not affect the recovery of the radioactive proteins and did not alter the sensitivity of the assay. This method is very simple and rapid and, since it is suitable for processing microcultures, permits detailed studies on the biology of small numbers of M phi.
D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D002448:Cell Adhesion; D004306:Dose-Response Relationship, Immunologic; D005260:Female; D006146:Guanidines / Q000494:pharmacology; D007158:Immunologic Techniques; D007930:Leucine / Q000378:metabolism*; D008264:Macrophages / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D014176:Protein Biosynthesis; D013154:Spleen / Q000166:cytology; D014238:Trichloroacetic Acid / Q000494:pharmacology
D000596:Amino Acids; D006146:Guanidines; D014238:Trichloroacetic Acid; D007930:Leucine