The human protein atlas blog


Validation of antibodies using GFP-tagged target

2015-12-15
Cell Atlas Co-localization HPA14 Immunocytochemistry Immunofluorescence Subcell Atlas Validation


Antibody HPA37362 targeting BOD1L1 overlaps with GFP-tagged target protein and the localization is identical to that of endogenous protein in non-transfected control cells

Version 14 of The Human Protein Atlas includes a new type of validation of antibodies that are used for determining the subcellular localization of a protein.

A set of antibodies have been analyzed in transgenic cell lines expressing GFP-tagged target protein at near-endogenous levels to confirm that the antibodies are capable of binding the target protein. The approved antibodies are then used to determine the subcellular localization of endogenous protein in a selection of cell lines. A high validation score is assigned to those genes where the same location(s) are observed for both tagged protein and protein detected using labelled antibody in non-transfected cells. With this approach the subcellular localization can be determined with high certainty.

This approach has been used for BOD1L1, a gene with no data on the subcellular location in UniProt. The antibody overlapped with tagged protein in transgenic cells and a nucleoplasmic location was observed with both methods.

The Human Protein Atlas currently contains GFP validation data for 100 genes and 123 antibodies.

Access all the antibody and antigen data for BOD1L1 here.

Also read our blog post on Increasing reliability by co-localization.