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Logo of biolprocBioMed CentralBiomed Central Web Sitesearchsubmit a manuscriptregisterthis articleBiological Procedures OnlineJournal Front Page
 
Biol Proced Online. 2012; 14: 4.
Published online Apr 2, 2012. doi:  10.1186/1480-9222-14-4
PMCID: PMC3388579
A microplate technique to simultaneously assay calcium accumulation in endoplasmic reticulum and SERCA release of inorganic phosphate
David C McMullen,corresponding author1 William S Kean,2 Ajay Verma,3 Jeffrey T Cole,3 and William D Watson3
1Department of Neurology, Uniformed Services University of the Health Sciences, B-3059, 4301 Jones Bridge Road, Bethesda, MD 20814, USA
2Department of Neurology, Uniformed Services University of the Health Sciences, B-3009, 4301 Jones Bridge Road, Bethesda, MD 20814, USA
3Department of Neurology, Uniformed Services University of the Health Sciences, B-3012, 4301 Jones Bridge Road, Bethesda, MD 20814, USA
corresponding authorCorresponding author.
David C McMullen: david.mcmullen.ctr/at/usuhs.mil; William S Kean: william.kean.ctr/at/usuhs.mil; Ajay Verma: Ajayverma401/at/gmail.com; Jeffrey T Cole: jeffrey.cole/at/usuhs.mil; William D Watson: wwatson/at/usuhs.mil
Received January 12, 2012; Accepted April 2, 2012.
Abstract
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.
Keywords: Calcium, SERCA activity, Microsomes, Inorganic phosphate, Malachite green
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