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Lysis with Ripa Buffer - too viscous, help needed

Lysis with Ripa Buffer - too viscous, help needed - Protocols and Methods Forum

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Old 10-27-2007, 06:40 PM
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Unhappy Lysis with Ripa Buffer - too viscous, help needed



Dear All,

I was doing a lysis of my cells in order to do a Western Blotting later on.

I had my cells in 6-well plates, highly confluent, and used Ripa Buffer with 1% SDS (I believe I was supposed to have used 0.1%)

After scrapping and centrifuging, I still had no pellet, and the solution was just as viscous as before. I repeated the centrifugation at 4C and still no pellet was formed.

I was wondering if you know what happened (for usually I get a normal pellet), and most importantly, how the samples could still be recuperated (they are frozen now).
They were from a very important experiment….

Thanks a lot!

Anne-Marie
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