Have you knocked out a gene and want to make sure that the corresponding protein is depleted? Whereas genotypic analyses can provide information regarding the percentage of CRISPR-edited sequences, protein analyses provide a more definitive assessment of knockout efficiency.

In this Tips and Tricks installment, we provide a primer on four commonly used protein assays: Western blot, immunocytochemistry, flow cytometry, and ELISA. The guide includes a summary and guidelines for each assay.

Synthego has created a Tips and Tricks series to provide useful information on how to enhance experimental techniques and protocols for CRISPR applications. Building your CRISPR skill set will enable you to take your genome editing experiments to the next level.

This Tips and Tricks edition provides a summary of four conventional protein analyses used for validating knockouts: Western blot, immunocytochemistry, flow cytometry, and ELISA. We include general guidelines as well as advice on how to optimize each assay.

The topics covered include:

  • Timing of protein analysis
  • Antibody optimization
  • Recommended controls
  • Example results

If you are planning a CRISPR knockout experiment and want to measure protein expression directly, this resource can help you choose the right assay and get started.