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

Biological Procedures Online

David C McMullen²  William S Kean¹  Ajay Verma³  Jeffrey T Cole³  William D Watson³ 

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

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

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

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

Others  :  793259
DOI  :  10.1186/1480-9222-14-4

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

【 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】

   
2012 McMullen et al; licensee BioMed Central Ltd.

【 图 表 】

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