Antibody Validation

Please note that we extensively validate antibodies applicable to RPPA analysis.

  • A single or dominant band on western blotting is required
  • Dynamic range and specificity is determined using:
    • Peptides, Phosphopeptides, Growth factors
    • Inhibitors, RNAi
    • Cells with wide levels of expression including 330 cell lines under multiple conditions on a single array
  • A Pearson correlation coefficient between RPPA and western blotting of greater than 0.7 is required
  • Reproducible results intra-slide or inter-slide are critical

Example of antibody validation of AKTS473:

Single band on western blot with dynamic changes by drug treatment

Relative protein concentration determined by RPPA Supercurvefitting and QC test, and the same set of samples run on RPPA

Pearson correlation between RPPA and western blotting greater than 0.7

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