Correct localization of proteins is critical for both the understanding and treatment of many diseases. High specificity immunoassays are of particular value for precision diagnostics use and in large-scale therapeutics development. The quality of these in situ protein detection assays depends mainly on the antibodies’ specificity against the target. Many commercial antibodies have low specificity, generating unspecific background staining.
The NaveniFlex proximity ligation platform enables the use of two primary antibodies against the target protein to generate a highly specific, quantifiable signal, contributing to improved sensitivity and noise reduction. Here we show a comparison of NaveniFlex and immunofluorescence (IF) using two antibodies to detect the ACE2 protein. The staining intensity from NaveniFlex is in excellent correlation with mRNA levels in both tissue samples (shown as TPM). The IF staining results in unspecific binding in lung tissue, leading to erroneous conclusions from localization studies using just one primary antibody.