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Immunoblot analysis

HY Hiroshi Yamaguchi
TO Toshiyuki Oda
MK Masahide Kikkawa
HT Hiroyuki Takeda
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Proteins were separated by SDS-PAGE in 5–20% gradient polyacrylamide gels (Nacalai Tesque) and transferred onto polyvinylidene difluoride (PVDF) membranes (Millipore). After blocking with 5% skim milk (Nacalai Tesque) in TBST (Tris-buffered saline containing 0.1% Tween-20), membranes were incubated with primary antibodies, followed by several washes and incubation with secondary antibodies (goat anti rabbit/mouse IgG antibody peroxidase conjugated; Sigma-Aldrich). Protein signals were visualized by ECL Select Western Blotting Detection Reagent (GE Healthcare) and observed using luminescent image analyzer (ImageQuant LAS4000mini; GE Healthcare).

Copyright and License information:  ©2018, Yamaguchi et al
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