Antibody validation

  • Standardized antibody validation methods
  • High purity, lot-to-lot consistency, and antibody reproducibility
  • Reliable antibody specificity and sensitivity

How does Miltenyi Biotec address the need for reproducible and validated antibodies?

With the introduction of recombinant antibodies in 2012, we made a significant investment into improving the quality and consistency of our antibodies (link to REA background page). The standardized antibody production process, starting from a defined DNA sequence, and the nature of recombinant antibodies ensure high purity and lot-to-lot consistency. 

In addition, recombinant antibodies do not display any undesired mixtures of heavy and light immunoglobulin chains, which is often the case with conventional hybridoma-derived antibodies⁵ (PMID: 29485921). Furthermore, our REAfinity™ Recombinant Antibodies have a mutated Fc region that abolishes any binding to Fcγ receptors, resulting in a background-free analysis. These advantages make REAfinity Recombinant Antibodies ideal tools for improving experimental reproducibility.

In 2020, we began the further step of providing antibody validation data directly on our product pages. We do this in order to make it even easier for our customers to choose the antibodies that best match their needs, and to decrease the validation efforts required on their researcher side. With well over 10,000 antibodies in our portfolio, this is an ongoing project and information is updated regularly, so please do check back from time to time. In addition, below you can find some insights on how we conduct our antibody validation process.

Extended validation stamp

All our antibodies are rigorously tested and validated before release. In the application section on the product page, you can find examples of typical performance data. In addition, we provide extended validation data highlighting details of antibody performance, specificity, and fixation compatibility. All antibodies for which any of these datasets are already available will be indicated with the extended validation stamp. That list will be continuously expanded, so be sure to check back soon if you don’t see what you need yet. 

Three pillars defining antibody quality and validation

REAfinity Recombinant Antibodies are based on three pillars of validation: reproducibility, specificity, and sensitivity. Please find more detailed information for each validation method below:

How to validate an antibody after purchasing?

Validation is of course not solely the responsibility of the supplier because antibody validation methods and antibody validation protocols are highly dependent on the intended application and experimental setup. For this reason, we recommend thoroughly researching the most suited validation methods for your application. Several articles have been published providing antibody validation guidelines, as well as publication guidelines. A selection is given below:

  • Uhlen et al. (2016) A proposal for validation of antibodies. Nat Methods,13(10):823-7 PMID: 27595404
  • STAR methods from CellPress
  • Lucas et al. (2020) MiSet RFC Standards: Defining a Universal Minimum Set of Standards Required for Reproducibility and Rigor in Research Flow Cytometry Experiments. Cytometry A, 97(2):148-155 PMID: 31769204
  • Roncador et al. (2016) The European antibody network's practical guide to finding and validating suitable antibodies for research. mAbs, 8(1), 27–36 PMID: 26418356