[Simultaneous assay of Ca(2+)-ATPase and Na+, K(+)-ATPase activities of osteoblast rat by malachite green colorimetic method].
The aim of this study was to find a method to measure simultaneously the activities of Ca(2+)-ATPase and Na+, K(+)-ATPase of rat osteoblast membrane. In this test, Ca(2+)-ATPase and Na+, K(+)-ATPase released inorganic phosphate (Pi) from ATP, and Pi was determined by colorimetric method using malachite green dye to show ATPase activities. Ca2+ concentration and osteoblast culture generation were important factors affecting ATPase activities. Ca2+ could elevate membrane ATPase activity. When Ca2+ concentration reached 40 mumol/L, ATPase activities were improved. Ca(2+)-ATPase activity decreased with the increase of osteoblast culture generation. Ca(2+)-ATPase activity was lower than Na+, K(+)-ATPase activity in osteoblast membrane. This method is simple, sensitive and reliable.
Cen
X
X
Department of Nutrition and Food Hygiene, School of Public Health, Chengdu.
Huang
Y
Y
Wang
R
R
Chen
Z
Z
Wu
Z
Z
D000818:Animals; D000252:Calcium-Transporting ATPases / Q000378:metabolism*; D002462:Cell Membrane / Q000201:enzymology; D002478:Cells, Cultured; D003124:Colorimetry; D004396:Coloring Agents; D005333:Fetus; D010006:Osteoblasts / Q000166:cytology / Q000201:enzymology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012394:Rosaniline Dyes; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism*
D004396:Coloring Agents; D012394:Rosaniline Dyes; C005095:malachite green; D000252:Calcium-Transporting ATPases; D000254:Sodium-Potassium-Exchanging ATPase