for western blot analysis
the buffer in which the antibody already is, written in the datasheet, or should I use already the bsa or milk solution that I use for final diluition?
Hello Silvia
Make aliquots of your freshly procured antibody in the buffer in which it is already in and store them in -20° C. Make aliquots in such a way that it is used for one time purpose. This will avoid repeat freeze-thaw. As you mentioned do not use BSA or skimmed milk.
Thank you.
hi
you should find optimal concentration of the antibody and then add to TBS-skim milk or TBS-BSA from company datasheet . ( optimal concentration between 0.5 ug/ml - 10 ug/ml )
Antibody staining >>>
There are two aspects here:
1. When a commercial antibody arrives you should aliquot it out into smaller volumes (ideally that required for a one time use, as suggested above) and store as directed in the provided datasheet. This does not involve adding any new buffer to the solution.
2. For incubating a western blot in primary antibody you should add the antibody into a fresh solution of whatever blocking solution you used. This is typically either milk or BSA made up in PBS-Tween.
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