Reporting research antibody use: how to increase experimental reproducibility

Helsby, Matthew A; Fenn, Joe R; Chalmers, Andrew D.

doi:10.12688/f1000research.2-153.v1

Cited by 21 publications

(12 citation statements)

References 7 publications

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“…There have been calls to standardize antibody production, characterization and use [ 24 , 25 ]. It is recommended that journals insist that authors be more transparent and consistent with reporting antibody details [ 26 ].…”

Section: Discussionmentioning

confidence: 99%

Authentication of collagen VI antibodies

2017

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BackgroundCollagen VI is a ubiquitously-expressed macromolecule that forms unique microfibrillar assemblies in the extracellular matrix. Mutations in the COL6A1, COL6A2 and COL6A3 genes result in congenital muscular dystrophy, arguing that collagen is critical for skeletal muscle development and function. Antibodies against collagen VI are important clinical and diagnostic tools in muscular dystrophy. They are used to confirm genetic findings by detecting abnormalities in the distribution, organization and overall levels of collagen VI in cells and tissues isolated from patients.MethodsMany antibodies have been raised against tissue-purified collagen VI and individual collagen VI chains, however few have been properly validated for sensitivity and chain specificity. To address this deficiency, we compared the ability of 23 commercially-available antibodies to detect extracellular collagen VI by immunohistochemistry on frozen tissue sections. To determine chain specificity, immunoblot analyses were conducted on cell lysates isolated from cells transfected with cDNAs for each individual chain and cells expressing all three chains together.ResultsOur analyses identified 15 antibodies that recognized tissue collagen VI by immunohistochemistry at varying intensities and 20 that successfully detected collagen VI by immunoblotting. Three antibodies failed to recognize collagen VI by either method under the conditions tested. All chain-specific antibodies that worked by immunoblotting specifically recognized their correct chain, and no other chains.ConclusionsThis series of side-by-side comparisons reveal at least two antibodies specific for each chain that work well for immunohistochemistry on frozen sections. This validation study expands the repertoire of antibodies available for muscular dystrophy studies caused by defects in collagen VI.Electronic supplementary materialThe online version of this article (doi:10.1186/s13104-017-2674-x) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 99%

Authentication of collagen VI antibodies

2017

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“…The application of antibodies across cancer biology—from basic research to diagnostics to therapeutics—is pervasive (13, 30). While standards have been applied, formally or informally, to the generation and use of diagnostic and therapeutic antibodies, basic research antibody reagents are generally devoid of any kind of consistency or standardization.…”

Section: Lack Of Standards In Basic Biological Researchmentioning

confidence: 99%

“…While standards have been applied, formally or informally, to the generation and use of diagnostic and therapeutic antibodies, basic research antibody reagents are generally devoid of any kind of consistency or standardization. Although blame is often pointed at the manufacturers, Helsby et al(30) contend that scientists and publishers are not doing everything they can to improve the situation and thus improve reproducibility, such as failing to provide thorough reporting of research antibody use or demonstrating that antibodies were validated. Moreover, publications routinely do not provide the host species, code number, and even the antibody supplier.…”

Section: Lack Of Standards In Basic Biological Researchmentioning

confidence: 99%

The Increasing Urgency for Standards in Basic Biologic Research

Freedman

Inglese

2014

Cancer Research

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Research advances build upon the validity and reproducibility of previously published data and findings. Yet irreproducibility in basic biological and preclinical research is pervasive in both academic and commercial settings. Lack of reproducibility has led to invalidated research breakthroughs, retracted papers, and aborted clinical trials. Concerns and requirements for transparent, reproducible, and translatable research are accelerated by the rapid growth of “post-publication peer review,” open access publishing, and data sharing that facilitate the identification of irreproducible data/studies; they are magnified by the explosion of high-throughput technologies, genomics, and other data-intensive disciplines. Collectively, these changes and challenges are decreasing the effectiveness of traditional research quality mechanisms and are contributing to unacceptable—and unsustainable—levels of irreproducibility. The global oncology and basic biological research communities can no longer tolerate or afford widespread irreproducible research. This article discusses (1) how irreproducibility in preclinical research can ultimately be traced to an absence of a unifying life science standards framework, and (2) makes an urgent case for the expanded development and use of consensus-based standards to both enhance reproducibility and drive innovations in cancer research.

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“…Simply put, suppliers cannot possibly generate all information by themselves, especially since an antibody is typically used for different applications and tissues. Furthermore, it was suggested that journals should add antibody reporting guidelines, instructing authors to include antibody reports 7 . The antibody search engine and review database pAbmAbs, ( http://pabmabs.com/wordpress/ ) presented by founder Dr Simon Glerup, also provides a useful platform for researchers to share antibody reports.…”

Section: Independent Antibody Validationmentioning

confidence: 99%

Working towards a consensus for antibody validation

2014

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Commercial research antibodies are the most commonly used product in the life science tools market, and their applications represent a significant investment of time and resources for researchers. Frequently however, the quality of antibodies does not meet the expectations of consumers, causing loss of valuable time and money. This can delay research efforts and scientific discovery, or even lead to false, irreproducible results to be published in the scientific literature. This raises the question of whether there should be universal standards for validating antibodies. During the 1 st International Antibody Validation Forum, hosted by St John’s Laboratory Ltd on October 15 th 2014 at Queen Mary University of London, scientists from academia and industry presented data highlighting quality issues arising from lack of antibody validation. While the forum identified significant current problems in the antibody market, it also discussed future opportunities for improved quality and transparency by encouraging data disclosure and data sharing. This article highlights the key issues and conclusions reached at the forum.

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Reporting research antibody use: how to increase experimental reproducibility

Cited by 21 publications

References 7 publications

Authentication of collagen VI antibodies

Authentication of collagen VI antibodies

The Increasing Urgency for Standards in Basic Biologic Research

Working towards a consensus for antibody validation

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