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Comparisons of ELISA and Western blot assays for detection of autophagy flux

By Sung-hee Oh, Yong-bok Choi, June-hyun Kim, Conrad C. Weihl and Jeong-sun Ju

Abstract

We analyzed autophagy/mitophagy flux in vitro (C2C12 myotubes) and in vivo (mouse skeletal muscle) following the treatments of autophagy inducers (starvation, rapamycin) and a mitophagy inducer (carbonyl cyanide m-chlorophenylhydrazone, CCCP) using two immunodetection methods, ELISA and Western blotting, and compared their working range, accuracy, and reliability. The ELISAs showed a broader working range than that of the LC3 Western blots (Table 1). Table 2 showed that data value distribution was tighter and the average standard error from the ELISA was much smaller than those of the Western blot, directly relating to the accuracy of the assay. Test-retest reliability analysis showed good reliability for three individual ELISAs (interclass correlation, ≥ 0.7), but poor reliability for three individual Western blots (interclass correlation, ≤ 0.4) (Table 3). Keywords: Autophagy, Mitophagy, ELISA, Western blot, Skeletal muscl

Topics: Computer applications to medicine. Medical informatics, R858-859.7, Science (General), Q1-390
Publisher: Elsevier
Year: 2017
DOI identifier: 10.1016/j.dib.2017.06.045
OAI identifier: oai:doaj.org/article:89a13db1078f44928502adbba10a112c
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