Fig. 3 | Nature Communications

Fig. 3

From: Enhanced validation of antibodies for research applications

Fig. 3

Validation of antibodies using capture MS. a Principle for validation of antibodies based on separation by SDS-PAGE and comparing the protein migration profile determined by an antibody (Western blot) and peptide identification performed by proteomics (capture MS). b Examples of orthogonal validation of two antibodies by capture MS in two different cell lines (RT4, U-251). The black arrow indicates theoretical molecular weight, and the blue bars represent number of peptides identified in each gel slice. c Summary of capture MS validation of antibodies (n = 2,888) showing if they were validated using other antibody validation pillars. d Distribution of theoretical molecular weights for the largest transcript of all (n = 19,628) human protein-coding genes divided into intracellular (blue) and secreted and membrane bound proteins (purple). Molecular weights are represented by the log2 value together with a molecular weight ladder from a typical Western blot assay used within the Human Protein Atlas program

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