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Scientists at Cell Signaling Technology (CST) use a combination of several approaches to validate antibodies in a number of applications, to provide you with the highest quality and most thoroughly tested reagents. The majority of our products are initially tested and approved by western blotting. If western analysis shows a single band of the appropriate molecular weight, the antibody may subsequently be tested and validated in other applications.

Western Blotting & Immunoprecipitation

All antibodies from CST are validated extensively by western blotting according to a rigorous validation protocol. Only if the antibody shows the appropriate banding pattern by western, will further validation be performed in other applications.


CST's antibodies are validated by our in-house IHC group in patient samples and clinically relevant mouse models of cancer. We currently offer over 500 antibodies validated for immunohistochemistry.


Validation for IF involves clearly demonstrating correct localization and outstanding performance for all IF-recommended antibodies. We have validated over 1,000 antibodies for immunofluorescence-based applications. Our IF group also routinely employs our antibodies on popular automated imaging and laser scanning high content platforms and can provide assistance in these applications.

Flow Cytometry

The flow cytometry group strives to provide the brightest signal and lowest background possible for all products we recommend in this application. CST offers over 600 antibodies recommended for flow cytometry, including a number of Alexa Fluor® and other conjugates optimized for this application.

Chromatin Immunoprecipitation

CST's ChIP-recommended antibodies have been validated in-house using the same rigorous standards as our other applications. Our antibodies and ChIP-grade magnetic beads are ideal for downstream ChIP–on-chip or ChIP-sequencing techniques as they provide high sensitivity and low background.


Cleavage Under Target & Release Using Nuclease (CUT&RUN) is a technology that can be used to explore protein-DNA interactions. It is an in vivo method that uses a target-specific primary antibody and a Protein A-Protein G-Micrococcal Nuclease (pAG-MNase) to isolate specific protein-DNA complexes1,2,3. Like all CST products, our CUT&RUN assay products are rigorously validated in-house to ensure that they will work in your experiments and generate data you trust.

ELISA Solutions

Cell Signaling Technology has a wide selection of sandwich antibody assays that let you measure important cellular signaling nodes and other cellular target proteins. In-house CST scientists identify optimal antibody pairs, develop, produce, and thoroughly validate our ELISA products to ensure the highest possible product quality, sensitivity, data consistency, and accuracy. Customer technical support is provided by the same scientists who develop our ELISA products.

  1. Skene PJ, et al. (2018) Nat. Protoc 13(5), 1006-1019.
  2. Meers MP, et al. (2019) BioRxiv 1, 569129.
  3. Skene PJ, and Henikoff S. (2017) Elife 6, e21865.