Knockout (KO) cell lines and antibody validation


Human knockout cell lines generated using CRISPR/Cas9 genome-editing.

In our recent article, we compared the advantages and disadvantages of different antibody validation techniques to help you choose the right method for your experiment. Though each technique highlights a unique type of validation, knockout validation gives the most precise answer to the question of specificity.

Abcam knockout validates its antibodies using the extensive library of knockout cell lines available from Horizon Discovery.  We are utilizing their human knockout cell lines generated from haploid cellular models using a CRISPR/Cas9 genome-editing system to bring you a range of premium knockout validated antibodies. This type of knockout cell line provides a complete loss-of-function phenotype from a single allele knockout and eliminates any masking of the knockout from a second allele seen in diploid cell models (1, 2, 3).

  • Human haploid cellular model
  • CRISPR/Cas9 genome-editing based knockout cell lines
  • Over 700 human knockout cell lines ready for testing Abcam's antibody portfolio
  • Constantly expanding library of KO cell lines

Fluorescent western blotting using anti-Cdk4 antibody [EPR4513] (ab108357) at 1/1000 dilution. Lane 1: Wild-type HAP1 cell lysate, lane 2:  Human Cdk4 Knockout cell lysate.  Target band at 34 kDa. Loading control band using anti-beta Actin (ab8227) at 42 kDa.


Extensive validation

Following our stringent quality control and validation processes, our antibodies are tested in both knockout and wild type samples using a western blotting based assay. We are in the process of expanding our knockout based antibody validation to other types of assays (i.e. ICC, IHC, Flow Cyt).



References

  1. Bürckstümmer T et al. A reversible gene trap collection empowers haploid genetics in human cells. Nat Methods. 2013. PubMed PMID: 24161985.
  2. Reiling JH et al. A haploid genetic screen identifies the major facilitator domain containing 2A (MFSD2A) transporter as a key mediator in the response to tunicamycin. Proc Natl Acad Sci U S A. 2011. PubMed PMID: 21677192.
  3. Carette JE et al. Haploid genetic screens in human cells identify host factors used by pathogens. Science. 2009. PubMed PMID: 19965467.
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