A simple method for incorporating aequorin into mammalian cells.
A simple method for incorporating aequorin into mammalian cells to measure cytosolic ionized Ca2+ is described and compared with scrape loading and hypoosmotic treatment (HOST). The procedure consists of incubating the cells for 10 min and centrifuging them at 200 g for 30 s in the presence of aequorin. This method incorporates the same amount of photoprotein as scrape loading but 70% less than HOST. Cytosolic ionized Ca2+ has been measured in hepatocytes, kidney cells and tubules, macrophages, and cardiac myocytes loaded with aequorin by this new procedure.
D000331:Aequorin; D000818:Animals; D002118:Calcium / Q000032:analysis; D002460:Cell Line; D002478:Cells, Cultured; D002498:Centrifugation; D003584:Cytological Techniques; D000882:Haplorhini; D066298:In Vitro Techniques; D007668:Kidney; D008164:Luminescent Proteins
D008164:Luminescent Proteins; D000331:Aequorin; D002118:Calcium