Fig 8. OneMinute®Advance is more efficient than Pierce¡¯s Restore and RestorePlus | |||
5% milk blocked Nitrocellulose Western Blot of 293 cell lysate
was probed with Mouse-anti-Human Actin (Sigma, A-5441) at 1:5000 dilution, HRP-Goat-anti-Mouse (Amersham) at 1:2000 dilution. The signal was detected using SuperSignal WestDura (Panel A). Blots were stripped with either
OneMinute®Advance Western Blot Stripping buffer ( 5min pretreatment + 1min stripping), or Pierce¡¯s Restore and Restore Plus (15 min stripping). Blots were re-probed with Mouse-anti-Human RIPK1 (BD, 610458) at 1: 200 dilution in 5% milk, HRP-Goat-anti-Mouse (Amersham) at 1: 2000 dilution. Signal was detected using SuperSignal WestDura (Panel B).
15min stripping with competitor Pirece¡¯s Restore and RestorePlus failed to remove high-affinity antibody to actin, and the stripping resulted in significant loss of RIPK1 protein. In contrast, OneMinute®Advance successfully stripped off high-affinity antibody to actin without loss of RIPK1 protein on membrane. |
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