KO Validated Antibodies

Figure 1: Schematic representation of how KO validated antibodies are created using CRISPR technology.

• We have created hundreds of CRISPR Knockout cell lines by introducing frame-shift mutations into the genomic DNA of target cells using sgRNA/CAS9.
  
  
• The CRISPR knockout cells are lysed and Western Blotting is performed using Wild-Type (WT) cells as a control.
  
  
• The Western Blots are then probed using our premium antibodies. Visualization of decreased protein expression is indicative of highly specific antibodies (Knockout Validated Antibodies).

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• KO validation is a means by which antibody specificity is confirmed by testing antibodies in a knockout (KO) cell line.
  
• Using KO cell lines (cells with no expression of target proteins) is the new gold standard to validate the specificity of antibodies.
  
  
• If a signal is detected from a KO cell line, then the antibody is not highly specific. This is due to the antibody binding to a similar protein within the sample. Antibodies with high specificity on the other hand will not produce any signal.
  
  
• KO validated antibodies must be shown to be specific, selective, and reproducible.

Figure 2: Immunohistochemistry of paraffin-embedded human colon using KO validated FASN antibody (CAB0462) at dilution of 1:100 (40x lens).

Figure 3: Immunofluorescence analysis of HeLa cells using KO validated KRT8 antibody (CAB1024) at dilution of 1:100. Blue: DAPI for nuclear staining.

• Antibodies are commonly used research tools. These proteins are used routinely in experiments such as Western blot, ELISA, immunostaining (immunohistochemistry (IHC) and immunocytochemistry (ICC)), immunofluorescence, and flow cytometry.
  
  
• For the findings of these experimental techniques to be reliable they must have an internally high standard of accuracy and precision.
  
  
• This is why there is a need for highly specific antibodies that can ensure reproducibility of results.