Deproteinizing Sample Preparation Kit - PCA

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  Functional Studies - Deproteinizing Sample Preparation Kit (ab284939)

  Functional studies - Deproteinization of serum sample.

  Different amounts of PCA were used to deproteinize 400 μL of serum following the kit protocol. Protein concentration remaining in solution OD280 (♦) and sample pH (•) was measured. Samples (neat serum ; serum filtered through a 10kD MW cutoff ; and PCA treated ) were diluted 20 times for convenience of measuring OD at 280 nm. A separate aliquot of the same PCA treated samples was mixed 1:1 with a pH 7.75 assay buffer and the pH of the mixture determined. The figure shows that sample/PCA ratio errors up to 20% can be tolerated with the resulting assay pH holding within approximately 0.5 pH units of the target assay pH. Using a 10 kDa MW cutoff filter removes approximately 98% of protein and PCA precipitation at a PCA/sample ratio of 1:4 removes approximately 95% of protein present.