Western Blotting, ELISA and MS

Cell Signaling/Transduction
1

hello everyone

Can you tell what key factors to consider when choosing between Western blotting, ELISA, and mass spectrometry for measuring phosphorylation levels of signaling proteins?

Regards

2

Hi Daniel

The choice b/w Western Blotting, ELISA and MS depends upon multiple factors:

  • Sensitivity: MS>ELISA>WB
  • Detection Limit: MS & ELISA (Picograms to Femtograms)> Western Blotting (ng)
  • Quantification: ELISA (easiest) >WB (semi) > MS (can be quantitative)
  • Cost: MS (Highest/ sample) > ELISA > WB (Lowest)
  • Reproducibility: ELISA > WB > MS

In addition to these, it is also important to note that, ELISA & WB are dependent upon Ab and thus are of limited value when you are trying to detect novel Phosphorylation sites, which can be circumvented by MS.

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