AAT Bioquest

What should I consider when I select antibodies?

Posted April 18, 2023


There are several factors to consider when selecting antibodies such as specificity, sensitivity, application, isotype, host species, reproducibility, and cost. Specificity is an important factor because one must ensure the antibody is specific to the target protein. To do this, the amino acid sequence of the protein being detected and cross-reactivity with other proteins should be considered. The isotype of the antibody is another factor to consider, as it may affect the specificity and sensitivity assays. The application the antibody is being used for is another factor (such as IHC, ELISA, western blot). The different types of applications may require the use of different types of antibodies such as polyclonal or monoclonal antibodies. Sensitivity of the antibody is another factor, as it should be sensitive enough to detect the target protein at the concentration used in the sample. Host species is another factor, as it may play a role in compatibility with the sample. As an example, an anti-fish antibody may not work with a fish sample. Reproducibility is another factor, and one should test the antibody under similar conditions in previous studies to verify it is high-quality. The last factor is cost, and one should note that cheaper antibodies may not give the best results and may require more optimization.

Additional resources

Physiology, Antibodies

Antibody and Protein Labeling

ReadiLink™ Rapid iFluor® 488 Antibody Labeling Kit *Microscale Optimized for Labeling 50 μg Antibody Per Reaction*