Yes. For overnight incubation use 4C. However, in my opinion, it would be better to block for an hour at RT, then incubate with the antibody overnight at 4C. That should give you a nice signal. You don't need to wash between blocking and ab, just a rinse will do, if its in the same buffer. Alternatively, carefully set your nitrocellulose face up on a piece of blotting paper for a few minutes, then sandwich between two clean dry pieces of blotting paper, wrap in aluminum foil, and set it in the -20 or -80 overnight. The cold will preserve your protein and keep the bands from spreading, and you can proceed with blocking and ab incubation the next day (or week!).
i linda,
definitely you can do it. We do it for some of the antibodies which gives non-specific signals.
What did you mean no signal specific , when can i said that this antibody gives no signal specific ...
Best regards
Yes. For overnight incubation use 4C. However, in my opinion, it would be better to block for an hour at RT, then incubate with the antibody overnight at 4C. That should give you a nice signal. You don't need to wash between blocking and ab, just a rinse will do, if its in the same buffer. Alternatively, carefully set your nitrocellulose face up on a piece of blotting paper for a few minutes, then sandwich between two clean dry pieces of blotting paper, wrap in aluminum foil, and set it in the -20 or -80 overnight. The cold will preserve your protein and keep the bands from spreading, and you can proceed with blocking and ab incubation the next day (or week!).
You can definitely do it.
In way to better preserve the BSA, I would suggest to do it at 4° C temperature. Anyway, as already mentioned, there is no need for such a long blocking time.
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