Comparison of Stain-Free gels with traditional immunoblot loading control methodology

Alexander Colella, Nusha Chegeni Farahani, Melinda Tea, Ian Gibbins, Keryn Williams, Timothy Chataway

    Research output: Contribution to journalArticle

    73 Citations (Scopus)

    Abstract

    Loading controls are necessary for semiquantitative Western blotting to compensate for loading errors. Loading control methods include the reprobing of membranes with an antibody against a constitutively expressed protein or staining the membrane with a total protein stain. We compared the loading control performance of recently released Stain-Free (SF) gels with Sypro Ruby (SR) and reprobing using β-actin. SF gels demonstrated superior performance in that they were faster, required fewer steps and consumables, and allowed the quality of electrophoresis and Western transfer to be assessed before committing to costly and time-consuming Western blots.

    Original languageEnglish
    Pages (from-to)108-110
    Number of pages3
    JournalAnalytical Biochemistry
    Volume430
    Issue number2
    DOIs
    Publication statusPublished - 15 Nov 2012

    Keywords

    • Immunoblot
    • Loading control
    • Semiquantitative
    • Stain-Free gel
    • Western blot

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