For the best experience on the Abcam website please upgrade to a modern browser such as Google Chrome

Hello. We're improving abcam.com and we'd welcome your feedback.

Hello. We're improving abcam.com and we'd welcome your feedback.

Infomation icon

We haven't added this to the BETA yet

New BETA website

New BETA website

Hello. We're improving abcam.com and we'd welcome your feedback.

Take a look at our BETA site and see what we’ve done so far.

Switch on our new BETA site

Now available

Search and browse selected products

  • A selection of primary antibodies

Purchase these through your usual distributor

In the coming months

  • Additional product types
  • Supporting content
  • Sign in to your account
  • Purchase online
United States
Your country/region is currently set to:

If incorrect, please enter your country/region into the box below, to view site information related to your country/region.

Call (888) 77-ABCAM (22226) or contact us
Need help? Contact us

  • My account
  • Sign out
Sign in or Register with us

Welcome

Sign in or

Don't have an account?

Register with us
My basket
Quick order
Abcam homepage

  • Research Products
    By product type
    Primary antibodies
    Secondary antibodies
    ELISA and Matched Antibody Pair Kits
    Cell and tissue imaging tools
    Cellular and biochemical assays
    Proteins and Peptides
    By product type
    Proteomics tools
    Agonists, activators, antagonists and inhibitors
    Cell lines and Lysates
    Multiplex miRNA assays
    Multiplex Assays
    By research area
    Cancer
    Cardiovascular
    Cell Biology
    Epigenetics
    Metabolism
    Developmental Biology
    By research area
    Immunology
    Microbiology
    Neuroscience
    Signal Transduction
    Stem Cells
  • Customized Products & Partnerships
    Customized Products & Partnerships

    Customized products and commercial partnerships to accelerate your diagnostic and therapeutic programs.

    Customized products

    Partner with us

  • Support
    Support hub

    Access advice and support for any research roadblock

    View support hub

    Protocols

    Your experiments laid out step by step

    View protocols

  • Events
    • Conference calendar
    • Cancer
    • Cardiovascular
    • Epigenetics & Nuclear signaling
    • Immunology
    • Neuroscience
    • Stem cells
    • Tradeshows
    • Scientific webinars
    Keep up to date with the latest events

    Full event breakdown with abstracts, speakers, registration and more

    View global event calendar

  • Pathways
    Cell signalling pathways

    View all pathways

    View all interactive pathways

Validating antibodies with knock-out technologies webinar

Related

  • Antibody validation with KO technology
    • Knock-out cell lines
      • Knock-out lysates
        • CRISPR-Cas9 technology: cell knockout generation
          • Custom CRISPR cell engineering services
            • Fast-track your research with KO cell lines
              • KO validation: Overcoming the perils of poor antibody specificity
                • Custom cell lines: Factors to consider before you begin

                  ​​​​How can knock-out validation support the reproducibility of your experiments?

                  ​Overview

                  • Watch Dr. Alejandra Solache discussing knock-out validation of our antibodies
                  • Why use knock-out validation?
                  • Abcam’s knock-out validation initiative
                  • About Dr. Alejandra Solache
                  • References

                  ​​Dr. Alejandra Solache, Vice President of New Product Development at Abcam, recently discussed how we validate our antibodies using knock-out (KO) technologies. Learn about the value of gene knock-out technologies in large-scale antibody screening by watching the video below.     

                  ​

                  ​You can see Dr. Alejandra Solache present more on antibody KO validation in this webinar from 21:03, which was a part of the “Antibody validation webinar series” hosted by the Antibody Society. For more webinars on Antibody Validation from experts in the field, you can also check out the Antibody Society's learning center.


                  Why use knock-out validation

                  Several studies have demonstrated the irreproducibility of life sciences reagents, which in the case of antibodies, frequently results from a lack of specificity to their targets of interest. This lack of specificity, related to issues such as cross-reactivity and off-target protein binding, has contributed to the so-called ‘reproducibility crisis’1,2.

                  KO validation is one of the most accepted and trusted validation processes for antibody specificity3,4. This robust technique can confirm the specificity of an antibody by testing it in a KO cell line, cell lysate, or tissue that does not express the target protein. A specific antibody should, therefore, produce no signal in the KO cell line but give a specific signal in the wild type sample. In this way, KO validation serves as a true negative control5.

                  Find out more about knock-out validation

                  Abcam’s knock-out validation initiative

                  To ensure that our antibodies are specific and provide reproducible results, we apply KO validation at a large scale. We validate our antibodies in multiple applications, including flow cytometry, western blot, and immunocytochemistry, using multiple antibodies per target. Antibodies that demonstrate specificity are given a special “KO validated” designation, whereas those exhibiting off-target binding are removed from our catalog. We have now confirmed the specificity of more than 2500 antibodies with KO validation for over 2000 protein targets.

                  Key learnings from Abcam’s KO validation initiative:

                  • Recombinant monoclonal antibodies, in general, demonstrate superior specificity and consistency compared to traditional monoclonal and polyclonal antibodies.
                  • A significant number of top-cited antibody clones have been observed to exhibit off-target effects.
                  • We recommend that scientists choose antibodies that have been validated in multiple applications, ideally with the use of KO technologies. Alternatively, you can validate an antibody yourself, using the appropriate KO cell line, KO cell lysate, or tissue.
                  • Confirming the specificity of an antibody can take time and effort, including the generation of a KO cell line using gene-editing technologies (on average 6 months to generate KOs). To help you accelerate your research and avoid tedious CRISPR engineering experiments, we now provide one of the largest collections of CRISPR-Cas9 KO lysates and KO cell lines.
                  • All our KO cell lines and lysates are reliable, ready-to-use, and allow you to confidently progress target identification and validation without the need to first establish a KO cell line. All our KO cell lines are validated by Sanger sequencing, and many have additional western blot data to confirm KO at the proteomic level.

                  About Dr. Alejandra Solache

                  Dr. Alejandra Solache joined Abcam in December 2013 and manages Abcam’s Global New Product Development and validation teams in Cambridge, UK, Hangzhou, China as well as Eugene and Branford, CT in the USA.  Alejandra has also been instrumental in developing Abcam’s innovation strategy and product pipeline. 

                  Before joining Abcam, Alejandra led the Antibody and Assay Development teams as R&D Director at EMD-Millipore, where she held various positions for eight years. She trained as a life science researcher where she gained expertise in immunology, cell signaling, and epigenetics through post-doctoral fellowships at the University of California, San Francisco, and the Trudeau Institute, Saranac Lake, NY. 

                  References

                  1. Bradbury, A., Plückthun, A. Reproducibility: Standardize antibodies used in research. Nature. 518, 27–29 (2015).
                  2. Baker, M. Reproducibility crisis: Blame it on the antibodies. Nature. 521, 274–276 (2015).
                  3. Madhusoodanan, J. Validating Antibodies: An Urgent Need. TheScientist. 2014.
                  4. Rhodes, K.J., Trimmer, J.S. Antibody-based validation of CNS ion channel drug targets. J Gen Physiol. 131, 407–413 (2008).
                  5. Bordeaux, J., Welsh, A., Agarwal, S., et al. Antibody validation. Biotechniques. 48, 197–209 (2010). 
                  Why use knockout validation?
                  Abcam’s knockout validation initiative
                  About Dr Alejandra Solache





                  Get resources and offers direct to your inbox Sign up
                  A-Z by research area
                  • Cancer
                  • Cardiovascular
                  • Cell biology
                  • Developmental biology
                  • Epigenetics & Nuclear signaling
                  • Immunology
                  • Metabolism
                  • Microbiology
                  • Neuroscience
                  • Signal transduction
                  • Stem cells
                  A-Z by product type
                  • Primary antibodies
                  • Secondary antibodies
                  • Biochemicals
                  • Isotype controls
                  • Flow cytometry multi-color selector
                  • Kits
                  • Loading controls
                  • Lysates
                  • Peptides
                  • Proteins
                  • Slides
                  • Tags and cell markers
                  • Tools & Reagents
                  Help & support
                  • Support
                  • Make an Inquiry
                  • Protocols & troubleshooting
                  • Placing an order
                  • RabMAb products
                  • Biochemical product FAQs
                  • Training
                  • Browse by Target
                  Company
                  • Corporate site
                  • Investor relations
                  • Company news
                  • Careers
                  • About us
                  • Blog
                  Events
                  • Tradeshows
                  • Conferences
                  International websites
                  • abcam.cn
                  • abcam.co.jp

                  Join with us

                  • LinkedIn
                  • facebook
                  • Twitter
                  • YouTube
                  • Terms of sale
                  • Website terms of use
                  • Cookie policy
                  • Privacy policy
                  • Legal
                  • Modern slavery statement
                  © 1998-2023 Abcam plc. All rights reserved.