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

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

Research output: Contribution to journalArticlepeer-review

4 Scopus citations

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).

Original languageEnglish
Pages (from-to)696-699
Number of pages4
JournalData in Brief
Volume13
DOIs
StatePublished - Aug 2017

Keywords

  • Autophagy
  • ELISA
  • Mitophagy
  • Skeletal muscle
  • Western blot

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