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IP beginner

IP beginner - Immunoprecipitation Forum

IP beginner - IP or Immunoprecipitation Forum.

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Old 03-01-2013, 02:55 PM
Pipette Filler
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I am a complete novice. I have extracted protein lysate from a tissue and have it a given concentration (3.452ug/ul). I have run a successful western blot loading 15ug of protein lysate (4.345ul) with lysate buffer and 2X reducing buffer.
I now wish to immunoprecipitate for a given protein of interest. How much should I start with before pre clearing with protein A beads? Most protocols I read refer to a volume of cell lysate with no info on actual quanitity of protein.

I was thinking of starting at 2mg so around 500ul of protein lysate? Does this roughly make sense?

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