In western blot, primary antibody usually incubates overnight at 4°C. But the signal after incubation for 1-2 hours at room temperature is also strong enough to detect. If I incubate at room temperature, can I reuse the antibody or throw it?
we use them multiple times and it has worked fine so far. We add roche block though, to help with conservation.
I reuse my antibodies reconstituted in buffer (milk, bsa, etc) for 1 month or roughly 6-8 times incubated even after doing it at room temperature. As long as you have sodium azide at 0.02% you will be fine with preventing bacterial growth in the buffer. After longer periods of time, milk for instance, starts to settle which can be difficult to get perfectly back into solution.
If you do not reuse your antibody you will be going through it very quickly which can get expensive. Also, it is nice to use the same working buffer with antibody if you are comparing multiple experiments.
I suspect the inclusion of a blocking agent is fairly critical to being to reuse the antibody. This ensure it does not stick non specifically to the container etc being used for storage.
- Badges
- Science topic
- Similar topics
- Computer Science and Engineering
- Bioinformatics