Selective assay of monomeric and filamentous actin in cell extracts, using inhibition of deoxyribonuclease I.
A simple and selective assay for monomeric and filamentous actin is presented, based on the inhibition of DNAase I by actin. In mixtures of monomeric and filamentous actin, only the monomeric form is measured as DNAase inhibitor. The total amount of actin in a sample can be determined after depolymerization of F actin with guanidine hydrochloride. The assay is rapid enough to detect changes in the polymerization state of actin in vitro over time intervals as short as 3 min. Data characterizing unpolymerized and filamentous actin pools in extracts of human platelets, lymphocytes and HeLa cells are presented.
D000199:Actins / Q000032:analysis* / Q000494:pharmacology; D001792:Blood Platelets / Q000032:analysis; D002118:Calcium / Q000494:pharmacology; D003851:Deoxyribonucleases / Q000037:antagonists & inhibitors*; D006146:Guanidines / Q000494:pharmacology; D006367:HeLa Cells / Q000032:analysis; D008214:Lymphocytes / Q000032:analysis; D046911:Macromolecular Substances; D008274:Magnesium / Q000494:pharmacology; D008970:Molecular Weight; D013329:Structure-Activity Relationship
D000199:Actins; D006146:Guanidines; D046911:Macromolecular Substances; D003851:Deoxyribonucleases; D008274:Magnesium; D002118:Calcium