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Western blotting (WB)

XT Xiaoli Tang
HL Huafei Lu
BR Bharat Ramratnam
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WB was performed as previously described21. Briefly, cell or tissue lysates were separated by 10% SDS–PAGE electrophoresis and electroblotted to nitrocellulose membrane (Bio-Rad). Blotted membranes were probed with their respective primary antibodies, rotating at 4 °C overnight. Membranes were washed three times in TBST buffer and probed with secondary antibody (IRDye800-conjugated Affinity Purified IgG) at room temperature for 1 h. Membranes were then washed three times in TBST buffer and direct infrared fluorescence detection was performed with a Licor Odyssey Infrared Imaging System21.

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