Hi there,
I am doing lot of western blotting lately for which I make fresh transfer buffer each time. I am thinking if the transfer buffer could be reused? If yes, how many times can we reuse the same?
I've never reused transfer buffers but out of curiosity about your question, I´ve performed a quick search and I find this article: Transfer buffer containing methanol can be reused at least five times and maintain a similar extent of protein transfer to PVDF membrane [Pettegrew et al. J Biomol Tech. 2009;20:93-5]. However, I think I could end up being contaminated with traces of protein, so perhaps it would be advisable only reuse it 2-3 times, it would have to be probed.
I have been reusing the transfer buffer. I just make sure that the color of buffer is not changed (turns yellow/orange). I think you can use it 2-4 times, if stored properly.
Hi Poonam Sagar ,
I use 3 times and after use i store the buffer at 4ºC. I also clean all material to avoid buffer contamination. Always worked for me.
Good luck,
Jacó
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