Fig 5. Minimal loss of transferred proteins was achieved by OneMinute®.

OneMinute: 10sec at RT, washing 3x 10 sec, No re-blocking.                    Competitor: 15min at 50℃, washing 3x 10min, re-blocking 1 hour, washing 3x 15min

5% milk blocked nitrocellulose Western blots of 293 cell lysate were stained with Ponceau S red (Panel A). Blots were stripped using either 2ml of OneMinute® Stripping Buffer (30 sec, RT) or 10ml of competitor stripping buffer (15min,  50℃), and then stained with Ponceau S red (Panel B). OneMintue® Buffer treated blot was washed 3x 10 seconds with TBST, and performed re-bloting directly without re-blocking. Competitor stripping buffer treated blot was washed 3x 10min, blocked with 5% milk for 1 hour, and washed 3x 15min with TBST. Blots were re-probed with Rabbit-anti-Human JNK1 (Santa Cruz, sc-747) at 1:200 dilution, HRP-Goat-anti-Rabbit (Amersham) at 1:10,000 dilution. The signal was detected using ECL substrate (Panel C).