Entering the Antibody Lottery: Tips for Avoiding Dubious Sources

Having reliable antibodies can make or break your assay. However, sourcing the right antibody for the job can sometimes feel like you’re entering the lottery; there are so many on the market, and often they just don’t live up to what was promised.

Cell Signaling Technology (CST) recognized this and have been working to develop and produce high-quality antibodies, as well as other kits and reagents. In addition, improving the quality and quantity of relevant data on antibody validation has been a high priority for almost two decades.

In March, CiteAb’s antibody validation initiative award recognized Cell Signaling Technology for the company’s efforts in proactively engaging a number of validation initiatives designed to improve the quality and quantity of data available to researchers.

To learn more about antibody validation and its importance, we got in touch with Anthony Couvillon, PhD, Scientific Marketing Project Manager at CST.

(Nb: As Anthony was extremely generous in sharing his insights, some responses have been shared as separate articles – see links below to full responses)

Michele Wilson (MW): The specificity of antibody binding is incredibly important for many research disciplines, yet it seems this is something that can’t always be relied on. Can you give us an idea of the extent of this problem?

This answer is covered in the article entitled: Antibodies in Research: The Good, the Bad, and the Validation Epidemic. Anthony provides insights on the sheer number of antibodies available for research, outlines the problems with many validation processes and discusses the properties that constitute a “good” or “bad” antibody.

MW: Are there any credibility “indicators” that can help people identify a trustworthy antibody supplier?

AC:

• Vendors that show no supporting data to match their claims regarding application performance, specificity or sensitivity. Only buy antibodies with data that supports the verification of the antibody in the application and species you intend to use.
• Avoid buying from vendors that don’t show validation data – a single band on a western blot is not validation, positive staining of a tissue sample is not validation. Validation data includes the use of a positive and negative model (cell lines, tissues, knockouts, etc.) to prove that the antibody is specific for the intended target.
• Look for recombinant monoclonal antibodies when you can – these are vastly more reproducible then their polyclonal counterparts, especially if you are engaging in a long-term project requiring a lot of antibody. Understanding that recombinant monoclonal antibodies only make up a small fraction of the available RUO antibodies, it is critical for the end user (and the vendor) to test each new lot of polyclonal antibody against the previous lot to ensure consistent results.
• Avoid vendors that don’t manufacture their own antibodies.  Too many antibody suppliers are merely resellers of other company’s products.  Resellers rarely do any of their own testing so either don’t show data or show data supplied by the original manufacturer. Google Image Search and some antibody search engines are great tools for weeding out resellers.
• Avoid companies that offer to send you another vial of the same antibody or credit your account instead of trying to help you solve the problem.  While this may sound like a good deal, a quality antibody manufacturer will provide scientific technical help BEFORE offering a refund or replacement.
• Product citations are a great source of information about how a specific product has been used, but the number of citations a product has should not be the sole basis for selecting an antibody. In some cases, a superior product with fewer citations might be available.
• Be wary of product reviews that sound too good to be true or have no supporting data to back up the claims.  Some third-party review sites can be very helpful, AntyBuddy and PAbmAbs are two of my favorites because they require users to supply data and protocol details supporting the product review, good or bad.
• If buying a commonly used clone sold by multiple vendors, for example the C-Myc (9E10) clone be sure to check the formulation (concentration) in addition to the price.  The cheapest option won’t always yield the most assays depending on the application and conditions you are using.

MW: What are the best approaches used to validate the specificity of antibody binding? How does the validation approach change with the technique in which the antibody will be used?

AC: There are antibody validation strategies that apply to all applications and others that are application-specific. It is critical to understand that each application must be verified independently and while validation across multiple applications can be supportive, verification of functionality and specificity in one application or model does not prove validation in another.

Read the full response here: Tips for Antibody Validation

MW: Can you tell us about the technical service you offer, and the level of collaboration you enter with customers when troubleshooting?

AC: We strive to provide our customers with a high level of technical support, working with them to ensure accurate, reproducible results. When a customer contacts CST Technical Support, they are speaking with scientists who are experts in the given assay and are extensively familiar with the products the customer is using. The scientists that validate, maintain and support our products are the ones taking the phone calls and answering the emails. We not only make protocol suggestions, but we collaborate closely with customers at all stages of their experiments, from assay design and protocol optimization to data analysis. This dedication has led to CST’s technical support team receiving numerous industry awards.

MW: This year, Cell Signaling Technology will celebrate its 20-year anniversary, and recently you have received awards for the “Life Science Reagent Supplier of the Year” and the “Antibody Validation Initiative.” What values and decisions have been key to your success as a business?

AC: Twenty years ago, CSTs founder, Dr. Michael Comb, realized that researchers needed a new, faster way to study how proteins and posttranslational modifications coordinated in response to cellular stimuli. This observation, in conjunction with the drive to perform high-quality research, led to the formation of CST and the production of the first posttranslational-specific rabbit antibodies. From the beginning, science and biology drove the business, not revenue or shareholders. To this day, CST is a private company, beholden to the scientific community and our mission of delivering the world’s highest quality antibodies and reagents in order to enable scientific discovery and therapeutic discoveries.

As science has evolved over the years, so have we. Adapting new strategies and technologies to maintain our position at the forefront of scientific research. Our close ties with the scientific community and relative size make it easy for CST to react quickly to the changing research landscape, focusing on those tools that will be the most impactful all while adhering to our strict quality principles. Dr. Comb’s guiding vision has remained the core of what makes Cell Signaling Technology a leader in the antibody industry – quality before quantity, science before business, discovery before acquiescence.

MW: Recently, CST have announced that they have formalized partnerships with eight local non-profit organizations (Change is Simple, Seeding Labs,  Greenbelt,  Ipswich River Watershed Association, Raw Art Works, Montserrat College of Art, Beverly Bootstraps, and LEAP for Education). This says a lot about your stance on corporate social responsibility! How did you go about selecting the organizations you have chosen, and how will they benefit from the partnership?

AC: CST has dedicated social responsibility as one of its four core values. Along with innovative science, best in class products and engaged employees, we believe it is our responsibility to partner with local organizations to give back to our community and to minimize our environmental impact.  The eight nonprofit organizations chosen as CST Corporate Partners align closely with our goals of exciting youth with science education, protecting our environment, reaching out to support the under served, and bringing our communities together through artistic expression.  These organizations are not new relationships to CST, in some cases we have provided financial and other support for many years. Every year, CST reaches out to community organizations, seeking to offer both financial and volunteers to support their ongoing initiatives.  These Partnerships will strengthen our commitment to by educating our employees about their great work internally, reaching out more than financially through our volunteer programs, and hosting them at our facilities for events like annual meetings or fundraisers.

Anthony Couvillon was speaking to Michele Wilson, Science Writer for Technology Networks.