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MAbs. 2016;8(1):27-36. doi: 10.1080/19420862.2015.1100787. Epub 2015 Sep 29.

The European antibody network's practical guide to finding and validating suitable antibodies for research.

Author information

1
a Monoclonal Antibody Unit; Spanish National Cancer Research Center ; Madrid , Spain.
2
b Immunology Unit, Department of Cell Biology; Immunology and Neurosciences ; Medical School, University of Barcelona ; Spain.
3
c NDCLS, Radcliffe Department of Medicine ; University of Oxford ; Oxford , UK.
4
d OxFabs; University of Oxford ; Oxford UK.
5
e Antibody and Vaccine Group; Cancer Sciences Unit; University of Southampton; Faculty of Medicine; General Hospital , Southampton , UK.
6
f Center for the Production of Diagnostic Reagents and for Research; Blood Transfusion Center of Slovenia ; Ljubljana , Slovenia.
7
g Center for Proteomics; Faculty of Medicine; University of Rijeka ; Rijeka , Croatia.
8
h Department of Immunology and Oncology ; Spanish National Center for Biotechnology; Consejo Superior de Investigaciones Científicas (CNB/CSIC) ; Madrid , Spain.
9
i Institute of Immunology; University Medical Center ; Hamburg , 20246 Germany.
10
j Custom Antibody Service (CAbS); IQAC-CSIC/CIBER-BBN ; Barcelona , Spain.
11
k Section of Immunobiology; Institute of Biomedicine; University of Leon ; Léon , Spain.
12
l Department of Tumorimmunology ; Radboud University Medical Center ; Nijmegen , The Netherlands.

Abstract

Antibodies are widely exploited as research/diagnostic tools and therapeutics. Despite providing exciting research opportunities, the multitude of available antibodies also offers a bewildering array of choice. Importantly, not all companies comply with the highest standards, and thus many reagents fail basic validation tests. The responsibility for antibodies being fit for purpose rests, surprisingly, with their user. This paper condenses the extensive experience of the European Monoclonal Antibody Network to help researchers identify antibodies specific for their target antigen. A stepwise strategy is provided for prioritising antibodies and making informed decisions regarding further essential validation requirements. Web-based antibody validation guides provide practical approaches for testing antibody activity and specificity. We aim to enable researchers with little or no prior experience of antibody characterization to understand how to determine the suitability of their antibody for its intended purpose, enabling both time and cost effective generation of high quality antibody-based data fit for publication.

KEYWORDS:

BLAST searches; CiteAb; EuroMAbNet; HLDA workshops; flow cytometry; gene silencing; immunohistochemistry; monoclonal antibodies; validation studies; western blotting

PMID:
26418356
PMCID:
PMC4966524
DOI:
10.1080/19420862.2015.1100787
[Indexed for MEDLINE]
Free PMC Article

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