Protein Precipitation Kit Protocol

Procedure:

  1. Add 1/20 volume Reagent A to sample in a centrifuge tube and mix well.
  2. Add 1/10 volume Reagent B to sample.
  3. Allow to precipitate for 20 minutes at room temperature. Precipitate is comprised of Reagent A:B complex along with trapped protein molecules.
  4. Collect precipitate by centrifugation and remove supernatant. The pellet will be large.
  5. Completely disperse pellet in acetone to dissolve away precipitated A:B complex. The solution should appear clear to cloudy, depending on protein concentration, with no visible clumps. Undispersed clumps will trap impurities which will be carried over into the final isolate.
  6. Collect proteins by centrifugation.
  7. To remove salts and surfactants, wash pellet with acetone, acetonitrile or 70% ethanol. This step may be repeated if desired for heavily contaminated samples, or for downstream applications requiring the highest purity proteins. Collect proteins by brief centrifugation if necessary.
  8. Redissolve pellet in desired buffer.
Catalog Number: 
EC-888

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