A microplate technique to simultaneously assay calcium accumulation in endoplasmic reticulum and SERCA release of inorganic phosphate

Biological Procedures Online

Methodology

William S Kean¹  William D Watson²  Jeffrey T Cole²  Ajay Verma²  David C McMullen³ 

[1] Department of Neurology, Uniformed Services University of the Health Sciences, B-3009, 4301 Jones Bridge Road, 20814, Bethesda, MD, USA;Department of Neurology, Uniformed Services University of the Health Sciences, B-3012, 4301 Jones Bridge Road, 20814, Bethesda, MD, USA;Department of Neurology, Uniformed Services University of the Health Sciences, B-3059, 4301 Jones Bridge Road, 20814, Bethesda, MD, USA;

关键词: Calcium;    SERCA activity;    Microsomes;    Inorganic phosphate;    Malachite green;   

DOI  :  10.1186/1480-9222-14-4

 received in 2012-01-12, accepted in 2012-04-02,  发布年份 2012

来源: Springer

【 摘 要 】

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.

【 授权许可】

CC BY   
© McMullen et al; licensee BioMed Central Ltd. 2012