Western DotBlot protocol?
crasmussen at anat.med.ualberta.ca
Tue Apr 25 00:54:02 EST 1995
blume at tigger.jvnc.net (John E. Blume) wrote:
>Does anyone have a favorite protocol for doing Western blots in some
>of dot/slot format?
>I have many samples to test for effect and I hope to avoid running a
>and transferring. Obviously, I am concerned about background and about
>membrane binding capacity, but it strikes me that someone must have
Offhand I can't remember the precise protein binding capacity of typical
membranes, but we have used nitrocellulose and Immobilon-P with great
success. We usually resuspend the protein of interest in either Milli-Q
water or PBS and apply to the filter. After dot/slot-blotting we then
proceed directly to the detection protocol. If you are worried about
overloading the filter, the simplest thing to do is apply a series of
serial dilutions. You will eventually find a linear response.
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