Antibody validation Practical guide to finding and validating suitable antibodies for research
Step 1. Define your initial requirements
The starting point when sourcing antibodies is to correctly identify your target protein. Having the complete information on the target antigen (approved nomenclature, protein sequence, splicing variants, post-translational modifications, proteolytic cleavage and sequence homology of the target protein) is key to determine how specific of your target protein can be your antibody. Unfortunately, and for different reason, this information is not always available. But if available, it is well worth performing some basic bioinformatics analysis of the target antigen to address some important questions:
- Is the immnunogen used unique for the target protein (sequence homology)?
- Can my antibody recognize different isoforms of the same protein?
- Can my antibody recognize other family members?
You should also evaluate which type of antibody may be the best reagent for your research, and for which application you are planning to use it.
To define your initial requirement you should: