De-risking Antibody Selection for Immunohistochemistry (IHC)

Scientists know that selecting good antibodies can often be a challenge. Many antibodies don’t work equally well for all applications (immunohistochemistry (IHC), immunofluorescence (IF), immunoprecipitation (IP), ELISA, Western blot etc.), and with a limited research budget it is important to select antibodies that are both specific to their target proteins and suitable for the desired experiment. There are currently over 300 antibody suppliers selling over 3 million antibodies, yet their successful use is highly dependent on the experimental context. Western blot validation data is common, but for other applications such as IHC it is often difficult to determine whether an antibody works well before you purchase.

Selecting good antibodies, particularly for IHC and immunofluorescence (IF), requires careful research. Several tools exist to help make informed decisions and minimize the experimental risk of selecting an antibody that does not work for your selected application. For example, there are a number of good antibody search engines that can streamline the process and filter antibodies based on application, reactivity, species, clonality etc. Search engine results, together with strong antibody validation data and purchase options such as trial size antibodies, help provide direction for a successful experiment. Despite all the available data, it is always advisable to perform an antibody optimization experiment to assess specificity and staining conditions prior to use on your experimental samples.

An alternative approach is to work with an established company that specializes in IHC such as Reveal Biosciences, as they offer a broad range of optimized assays and have experienced scientists available who can help select new antibodies as necessary for your target of interest.

Antibody Search Engines/Databases

Antibody search engines driven by artificial intelligence (AI) are becoming popular ways to identify antibodies for a specific application. These search engines help de-risk your antibody selection by providing references to publications containing data and methods from scientists who have used these products before. Several of our favorite antibody search engines are listed below:

CiteAb – is a data focused antibody search engine that allows researchers to search over 3,894,038 antibodies, with 1,629,990 citations from 157 suppliers based on published literature. Antibodies are ranked based on the number of citations, and each paper is referenced, allowing scientists access to the methods used in each case. This website offers the advantage of enabling researchers to perform antibody searches based on application and species reactivity that are not only reported by the antibody vendor, but also those that have alternative published uses.

BenchSci – uses AI to extract antibody information from millions of scientific papers, which can then be searched, based on a wide range of parameters including application, species, and reactivity. BenchSci also provides over 3 million images from publications, vendors and independent organizations.

Biocompare – The Biocompare Antibody Search Tool lets researchers search over 3 million antibodies from hundreds of antibody suppliers quickly and easily. Their antibody database can be searched by target antigen, species reactivity, host, conjugate or application.

Antibodypedia – Antibodypedia is an open access, online antibody database that uses an evidence-based scoring system to rank each antibody. The Human Protein Atlas includes an Antibodypedia link for each marker, allowing filtering by validation method, application, host, and reactivity. Antibodies are listed by vendor with scoring by application.

Trial Size Antibodies

Many companies including Cell Signaling Technologies, Abcam, Novus Biologicals and R&D Systems provide trial size antibodies at a reduced price, allowing scientists to test the product in their application of interest before committing to purchase an antibody at full price. This approach can save money, but also allows the option of testing antibodies from multiple vendors to select which will perform best in your experiment. Keep in mind that the antibody industry includes a large network of distributors, so it is common for multiple companies to sell the same antibody or clone under different branding. Pricing can vary significantly and it is usually preferable to purchase from the original manufacturer if possible.

Antibody Validation Data

Antibody companies are increasingly providing comprehensive product validation data and client reviews on their web sites in order to increase the researcher’s confidence in their products. Abcam does this particularly well, with a good range of reviews available for most of their products. Incentives exist for researchers reviewing new antibodies or those used for new applications. Antibody companies that provide validation data often see increases in sales, easily justifying the cost of the initial testing. The Human Protein Atlas is also a good resource when selecting antibodies as IHC images are available for most of the proteins in the database, plus associated links to Antibodypedia.

IHC Services

Reveal Biosciences offers over 250 optimized IHC antibody protocols available as part of our comprehensive IHC services. Our scientists have carefully selected and optimized the staining conditions for each antibody using automated Leica Bond immunostainers. This automated approach produces consistent, reproducible results using antibodies that have already been tested for IHC. We also offer whole slide imaging and image analysis to generate quantitative data on a cell-by-cell basis from your slides.

High Quality Antibodies Result in Better Data: CD3-CD8 2-plex Immunochemistry Performed at Reveal Biosciences