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5% milk blocked nitrocellulose Western blots of 293 cell
lysate stimulated with or without Sorbitol (0.6M, 20min)
were probed with Rabbit-anti-Human phospoho-JNK1/2
(T183/Y185) (Cell Signaling Technology, #9251) at 1:1000
dilution, HRP- Goat-anti-Rabbit (Amersham) at 1:10,000
dilution. The signal was detected using ECL substrate
(Panel A). Blots were stripped using 2ml of OneMinute®
Stripping buffer (30 sec, RT), and washed 3x 10 seconds
with TBST (Panel B). Blots were re-probed with Mouse
anti-Human JNK1/2 (BD, #554285) at 1:1000 dilution, HRP-Goat-anti-Mouse
(Amersham) at 1:2,000 dilution and
detected with ECL substrate (Panel C).
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