western blotting transfer buffer - re-usage possible? (Apr/04/2008 )

hallo,

how often western blotting transfer buffer (e.g. towbin) could be re-used.

thanks

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I think you can use it 2-4 times (at least I do that). However, it also depends how long you store your buffer (1-2 weeks in a dark bottle at 4 degree). If the color has changed (for example yellowish), then you should make a fresh one of course.

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Use it once only. Don't risk having to repeat the entire experiment because of poor protein transfer.

If MeOH is becoming a major expense, you can switch to a dry transfer apparatus.

peace

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you can use it 2- to 3-times then it becomes undefined particularly the amount of methanol...

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The number of times we can use transfer buffer depends on voltage you use for transfer.
Generally if it is overnight transfer we use the buffer for 5 times atleast. If the transfers are 3 hour transfers then we use buffer 3 times.

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Well, we use it like 10 times without problems. At the fifth time, we discard 50% of it and re fill the bottle with new buffer

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