The absence of actin antibody after stripping by OneMinute®Advance shows the primary antibody has been totally stripped off.
| Fig 10. OneMinute®Plus is a bit better than OneMinute® stripping buffer, and OneMinute®Advance has the highest stripping efficiency. | |||
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5% milk blocked Nitrocellulose Western Blot of 293 cell lysate was probed with actin antibody (Mouse-anti-Human Actin from Sigma, A-5441) at 1:5000 dilution, and then 2nd antibody
(HRP-Goat-anti-Mouse from Amersham) at 1:2000 dilution. The signal was detected by SuperSignal WestDura (Panel A). Blots were stripped with
OneMinute® Stripping Buffer (30 sec), or
OneMinute®Plus Stripping Buffer (30 sec), or PBST (1xPBS with 0.1% Tween 20) as a negative control (-) for 30 sec, or
OneMinute®Advance Stripping Buffer (5 min pretreatment + 30 sec stripping). The blots were then re-probed with 2nd antibody
(HRP-Goat-anti-Mouse from Amersham) at 1:2000 dilution to test the presence of the actin antibody.
The absence of actin antibody after stripping by OneMinute®Advance shows the primary antibody has been totally stripped off. |
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