Reuse antibodies for further incubation in Western blot - (Mar/16/2005 )

I have a question on the usage of antibodies in general for western blotting. How many times can one reuse the primary and secondary antibodies?

Please share from your experiences.

Many thanks

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I reuse them as long as the signal stays good. You can't reuse is though if you incubate with BSA or milk.

Simon

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I reuse it as long as the signal stays good even when I incubate with 1% BSA or milk. I keep it at 4 degrees fridge with 0.02% Na Azide.
and it works fine, real fine

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hi
i reuse antibodies until the signal becomes reared. But if the milk stinks, i waste the solution.

is that product to prevent the milk degradation?
i use to store my antibodies solutions at -20°....but if i add Na azide should i store at 4 or -20 is still fine?

Thanks in advance.

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Hi
Na Azide prevents bacterial growth, so it won't stink for longer time
use it at 0.02% it works fine

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I store my Ab's diluted in milk - I find it is better to dissolve the milk powder in autoclaved/sterilised PBS before adding the antibody too - this helps it keep for longer at 4 degrees.

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