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Measurement of ghrelin concentration

CI Chika Ikenoya
ST Shota Takemi
AK Arisa Kaminoda
SA Sayaka Aizawa
SO Shiomi Ojima
ZG Zhi Gong
RC Rakhi Chacrabati
DK Daisuke Kondo
RW Reiko Wada
TT Toru Tanaka
ST Sachiko Tsuda
TS Takafumi Sakai
IS Ichiro Sakata
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Plasma and medium concentrations of acyl-ghrelin and des-acyl ghrelin were measured by an acylated Ghrelin (mouse, rat) Express EIA Kit (A05117, SPI Bio, Montigny-le-Bretonneux, France) and an Unacylated Ghrelin (mouse, rat) Express EIA Kit (A05118, SPI Bio). Acyl-ghrelin and des-acyl ghrelin concentrations were determined according to the manufacturer’s protocol. For acyl-ghrelin ELISA, the intra-assay coefficient of variation = 5.7%, and the inter-assay coefficient of variation = 6.1%. For des-acyl ghrelin ELISA, the intra-assay coefficient of variation = 5.2%, and the inter-assay coefficient of variation = 5.5%. Absorbance data were collected by using a Bio-Rad iMark Microplate Reader spectrophotometer, using Microplate Manager 6 Software (Bio-Rad, Hercules, CA, USA). Total ghrelin was calculated by adding acyl-ghrelin and des-acyl ghrelin concentrations together.

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