Department of Neurology, Uniformed Services University of the Health Sciences, B-3059, 4301 Jones Bridge Road, Bethesda, MD 20814, USA.
Biol Proced Online. 2012 Apr 2;14(1):4. doi: 10.1186/1480-9222-14-4.
Traditional analyses of calcium homeostasis have separately quantified either calcium accumulation or release mechanisms. To define the system as a whole, however, requires multiple experimental techniques to examine both accumulation and release. Here we describe a technique that couples the simultaneous quantification of radio-labeled calcium accumulation in endoplasmic reticulum (ER) microsomes with the release of inorganic phosphate (Pi) by the hydrolytic activity of sarco-endoplasmic reticulum calcium ATPase (SERCA) all in the convenience of a 96-well format.
传统的钙稳态分析分别对钙积累或释放机制进行了量化。然而,要定义整个系统,需要多种实验技术来同时检测积累和释放。在这里,我们描述了一种技术,该技术将内质网(ER)微粒体中放射性标记钙积累的同时定量与肌浆内质网钙 ATP 酶(SERCA)水解活性导致的无机磷酸盐(Pi)释放相结合,所有这些都在 96 孔格式的便利性下进行。