Tuesday, 01 July, 2008 | Supplied by: Merck

Prestige Antibodies, powered by Atlas Antibodies, are developed and validated by the Human Proteome Resource (HPR) project. The Human Protein Atlas (HPA) uses an antibody-based proteomics approach to test all the antibodies to ensure specificity. This approach combines the high-throughput generation of monospecific antibodies with protein profiling in standard sets of tissue and cell microarrays.

Each antibody produced begins with the selection of a protein epitope signature tag (PrEST). The PrEST fragment is typically 50-150 amino acids in length and is as dissimilar as possible to other proteins. The recombinant PrEST protein is utilised as the antigen in generating affinity purified monospecific antibodies.

Plasmid inserts are sequenced to ensure that the correct PrEST sequence is cloned. The size of the resulting recombinant protein is analysed using mass spectrometry to ensure that the correct antigen has been produced and purified.

To ensure antigen specificity, the antibodies are tested on protein arrays spotted with PrEST fragments. The antibodies are then subjected to testing using a universal immunohistochemistry (IHC) protocol against hundreds of normal and disease tissues. This protocol allows researchers to directly analyse samples without needing to optimise the antibody for different tissues.

Additionally, the antibodies are further tested by Western blot analysis and immunofluorescent microscopy.


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