Antibody anarchy: A call to order

Metabotropic glutamate receptor 3 (mGlu3; mGluR3; GRM3) in schizophrenia: Antibody characterisation and a semi-quantitative western blot study

“…Notably, this latter signal appeared plausible, with enhanced staining around the cell membrane, consistent with the membrane enrichment seen in the western blots. These observations highlight the need for caution and for additional immunohistochemical controls even for antibodies validated by immunoblotting (Holmseth et al, 2006, Rhodes and Trimmer, 2006, Baker, 2015). We did not pursue more detailed strategies, such as pepsinisation, or antigen retrieval, which might have permitted different and more valid immunohistochemical findings, as was reported for anti-mGlu2 antibodies (Lorincz and Nusser, 2008).…”

“…These issues affect receptors of interest in schizophrenia research, including mGlu1 receptors (Ayala et al, 2012) and muscarinic receptors (Pradidarcheep et al, 2008) just as much as other target proteins. Reflecting these concerns, there are calls for more careful, multifaceted, and method-specific validation of antibodies (Holmseth et al, 2006, Lorincz and Nusser, 2008, Bordeaux et al, 2010, Baker, 2015). Amongst other considerations, appropriate characterisation should include demonstration that the antibody binds specifically and selectively to band(s) of the predicted molecular weight of the target protein when present, and with absence of signal when the target protein is absent.…”

Challenges in Developing a Biochip for Intact Histamine Using Commercial Antibodies

“…Several authors have addressed the issue and claimed for better characterization and profound information on the specificity and cross-reactivity of the antibodies [19][20][21][22][23][24][25][26][27][28]. The antibodies in the market are often insufficiently characterized and furthermore they might possess batch-to-batch variation, usually in much larger extent with polyclonal antibodies compared to monoclonal.…”

“…The recommended applications are mainly immunohistochemistry (IHC) or immunocytochemistry (ICC), but also western blot, EIA and ELISA. Recently, several studies have pointed out the quality issues and traceability problem concerning commercial antibodies and the lack of quality in the antibody market has been widely recognized by the scientific community [19][20][21][22][23][24][25][26][27][28]. Often, the characterization and specifications provided by the manufacturers are somewhat incomplete and, furthermore, the authors often fail to identify the exact antibody used [28][29][30] for other researchers to be able to purchase the same product.…”

RRIDs: A Simple Step toward Improving Reproducibility through Rigor and Transparency of Experimental Methods

“…The volatility of the commercial market is one reason why groups that are coming together to set standards about what constitutes a validated antibody are recommending that in addition to specifying information that will need to be included in the material data sheet and what sorts of experiments need to be done to verify that an antibody is working within a researcher’s lab (IWGAV and GBSI; Baker 2015b; Alm et al, 2016), each antibody should be registered with the Antibody Registry and obtain a stable identifier.…”