LibGuides: Resources for Research Rigor &amp; Transparency: Antibody & Cell Line Validation

For the context within which they will be used, antibodies must be shown to be specific, selective, and reproducible. Antibodies should be chosen based on these criteria and be additionally validated with appropriate controls in the course of research. The resources below include information about antibody validation techniques, but also resources to identify antibodies previously validated for specific uses that may help in initial selection.

Antibodypedia
Almost 2 million antibodies. Search and compare from different vendors based on data from evaluation of the reagents in different techniques.
Antibodies Online
Independently validated antibodies get called out. Less than half of those tested were successfully validated.
JCN Antibody Database
Antibodies used for immunohistochemistry by papers published in Journal of Comparative Neurology from 2006 to present. With Antibody Registry numbers and PubMed IDs.
Antibody Comparison Web Sites (from Pivotal Scientific)
These sites often provide information on antibody validation for specific applications/species, which may also need to be reported.
CiteAb
Database that ranks antibodies by the number of times they've been cited in publications. See database paper at Biomed Central.
Immune Epitope Database and Analysis Resource
Repository of data about epitope recognition
Antibody Validation
In this review, we highlight the common pitfalls when working with antibodies, common practices for validating antibodies, and levels of commercial antibody validation for seven vendors. Finally, we share our algorithm for antibody validation for immunohistochemistry and quantitative immunofluorescence.
A proposal for validation of antibodies (Nature Methods)
"We convened an ad hoc International Working Group for Antibody Validation in order to formulate the best approaches for validating antibodies used in common research applications and to provide guidelines that ensure antibody reproducibility. We recommend five conceptual 'pillars' for antibody validation to be used in an application-specific manner."
Improving Reproducibility: Best Practices for Antibodies
Standardize Antibodies used in Research by Andrew Bradbury and Andreas Plückthun

Validation of cell line's species, sex, and tissue of origin are essential to ensuring that results can be interpreted correctly. Journals may already ask you to report the source and authentication status of cell lines. For example, information on what Nature is looking for can be found here.

First step (and simplest): check your cell line(s) against the list of known cross-contaminated or misidentified cell lines available on the NCBI Biosample home page.

NCBI Biosample - Browse Known Misidentified Cell Lines

The NIH (and more frequently, journals as well) will want to know how you plan to authenticate your cell lines. Generally, STR testing and mycoplasma contamination testing are recommended. In both tests, the method used, the outcome, and the date last authenticated should be supplied. The materials linked to below contain more information on authentication best practices and standards.

Guidelines for the use of cell lines in biomedical research
General guidelines, but contains specific sections on authentication and contamination.
Standards for Cell Line Authentication and Beyond
Different genomic technologies have been applied to cell line authentication, but only one method (short tandem repeat [STR] profiling) has been the subject of a comprehensive and definitive standard (ASN-0002). Here we discuss the power of this document and why standards such as this are so critical for establishing the consensus technical criteria and practices that can enable progress in the fields of research that use cell lines. We also examine other methods that could be used for authentication and discuss how a combination of methods could be used in a holistic fashion to assess various critical aspects of the quality of cell lines.
ANSI Standard - Authentication of Human Cell Lines: Standardization of STR Profiling
This document elaborates a standardized procedure for unambiguous authentication and identification of human cell lines using STR profiling. The overall objective of this standard is the specification of the methodology for DNA extraction and for STR profiling, data analyses, quality control of the data, interpretation of results, and implementation of a searchable public database

Additional help and resources:

ATCC STR Database
ATCC has developed a comprehensive database of short tandem repeat (STR) DNA profiles for all of our human cell lines.
Cell-line authentication demystified (Marx, Nature Methods 2014)
Cellosaurus
Knowledge resource on cell lines. Includes much information on each, including STR profiles
International Cell Line Authentication Committee (ICLAC)
JCRB Cell Bank
A Resource for Cell Line Authentication, Annotation and Quality Control (Uhlen et al., Nature Methods 2016)
Recommendation of Short Tandem Repeat Profiling for Authenticating Human Cell Lines, Stem Cells, and Tissues (Barallon et al., In Vitro Cell Dev Biol Anim. 2020)
Know Thy Cells: Improving Biomedical Research Reproducibility by Leonard P. Freedman
Changing the Culture of Cell Culture: Applying Best Practices and Authentication to Ensure Scientific Reproducibility by Leonard P. Freeman, Mark C. Gibson and Richard M. Neve
Authentication for non-human cell lines from ICLAC
Mouse cell line authentication