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...Read more


Increasing reliability by co-localization

2015-11-10
Cell Atlas Co-localization HPA14 Immunocytochemistry Immunofluorescence Subcell Atlas Validation

Subcellular localization obtained using IF (left) and GFP (FP, right) for the endoplasmic protein FKBP7 (top) and the FHL2 protein localized to focal adhesion sites (bottom).

Immunofluorescence and fluorescent-protein tagging show high correlation for protein localization in mammalian cells

The Human Protein Atlas applies antibodies for a variety of applications to map protein expression in different tissues and also at the subcellular level. Within the subcellular protein atlas, immunofluorescence (IF) is used to uncover the localization of proteins to different organelles. To ensure an accurate localization of each and very protein, the antibodies have to be specific to their target protein...Read more