Antibody validation is a critical step in the journey towards obtaining consistent reproducibility in science. To ensure they are both specific and sensitive, we validate our antibodies through a variety of methods including:
 
  • Testing on multiple cell and tissue types with a variety of known expression levels.
  • Validation in multiple applications as a cross-check for specificity and to provide additional clarity for researchers.
  • Comparison to existing antibody clones.
  • Using cell treatments to modulate target expression, such as phosphatase treatment to ensure phospho-antibody specificity.

Testing on Multiple Cell Lines

Western blot of whole cell extracts from both human and mouse cell lines known to have a range of expression of PIK3R2 (clone W17020A).

Testing in Multiple Applications



IHC staining of FFPE human brain tissue (top). ICC staining of Sh-SY5Y cells (bottom). Tissue and cells were stained anti-Tubulin β3 (clone TUJ1).


Western blot of brain lysates from human (lane 2), mouse (lane 3), and rat (lane 4) probed with anti-Tubulin β3 (clone TUJ1).

One of the most trusted ways to ensure specificity is to use knockdown or knockout models. This is especially true for ubiquitously expressed proteins where there aren’t negative control or low expressing cell types available. In these cases, we also use either siRNA-mediated knockdown or CRISPR/Cas9-mediated knockout of the target protein.

Western blot of total lysates from HeLa and HeLa AKT1CRISPR/Cas9 knockout (KO) cells probed with anti-AKT1 (clone O94E10).

Check out our updated our Stem Cells and Development wall poster incorporating the latest in stem cell biology including development into the three major germ layers, induced pluripotent stem cells, and how the niche affects stem cell development. Visit our Stem Cell Poster Webpage to view products to study stem cells, download the poster, or request a physical copy to hang in the lab.

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