Many epigenetics techniques and projects rely on high-quality antibodies, but not all commercial antibodies perform well enough to get the job done. On top of that, there’s no uniform standard, or easy steps to fully validate a good antibody. With help of the Park lab at Harvard Medical School, there is now a place that can help separate the good antibodies from the not-so-good ones, called The Antibody Validation Database, and it was designed to “collect and to share experimental results on antibodies that would otherwise remain in individual laboratories, thus aiding researchers in selection and validation of antibodies.”
The site began last year with 200+ histone antibodies tested as part of the ENCODE (Encyclopedia of DNA Elements) and Roadmap Epigenomics projects at the NIH. The initial results were published in Nature Structural & Molecular Biology, (Egelhofer et al. December 2010) where they tested antibodies to 57 different histone variations. Here’s the criteria that was used:
- Western blot: Pass if the histone band constituted at least 50% of the total nuclear signal, was at least ten-fold more intense than any other single nuclear band, and was at least ten-fold more intense than recombinant, unmodified histone.
- Dot blot: Pass if at least 75% of the total signal was specific to the cognate peptide.
- ChIP-chip/seq: Pass if the Pearson correlation is more than 0.8 on any pair of ChIPs performed from independent preparations matched for stage, cell type or biological tissue
After finding that over a quarter of the antibodies failed their validation process, the team now recommends that a similar validation be done on antibodies before they go into an experiment…or at least save yourself some trouble and check their database before you get started. Check out some of the antibodies sources that have been tested so far:
- Active Motif
- Hiroshi Kimura Lab
- Lake Placid
- LP Bio
- Santa Cruz Biotech
- Thomas Jenuwein Lab
The Antibody Validation Database is still a work in progress though, and there are still many more antibodies to be tested, so they welcome involvement from the rest of the scientific community, like these labs that have already joined in:
- Bing Ren Lab
- Gary Karpen Lab
- Julie Ahringer Lab
- Mitzi Kuroda Lab
- Sarah Elgin Lab
- Strome Lab
- Jason Lieb Lab
- Vincenzo Pirrotta Lab
Find out more about the antibodies that might be sitting on our shelf at the Antibody Validation Database.