What is KO cell lysate?
Gene knockout cell lysates are the cell homogenate in RIPA buffer made with double-knockout cell lines. The Gene knockout cell lines developed by CRISPR. The protein concentration was determined with BCA assay. A vial of lysate from the parental cell line was also included as an internal control. To facilitate transportation and protein, the products are supplied as lyophilized proteins or SDS format.
What are the applications of the KO cell lysate?
Validating* an antibody’s specificity is crucial to ensuring the absence of nonspecific binding and, therefore, the highest level of functionality. Testing antibody performance against genetically modified samples is one way to verify that an antibody recognizes a specific target. EdiGene’s KO cell lysate is immediate serve as negative controls for western blots and ultimately to validate the specificity of antibodies as for new antibodies validation standard (antibody KO validation).
We are expecting that novel applications such as a negative control for functional assay. Feel free to contact us if you have any ideas on how to use this material.
What is supplied in each KO lysate products?
Each KO cell lysate kit includes 2 vials:
· KO Cell lysate (100ug)
· Parental Cell lysate (100ug)
*The use or any variation of the word “validation” refers only to research use antibodies that were subject to functional testing to confirm that the antibody can be used with the research techniques indicated. It does not ensure that the product(s) was validated for clinical or diagnostic uses.
For Research Use Only. Not for use in diagnostic procedures.
- Case Study
In the example below, validate the specific of Origene’s MET monoclonal antibody (SKU. UM800082) using our MET knockout cell lysate (SKU. L0018998301) in western blot. The result showed significant disappear in protein expression, confirming specificity of the anti-MET antibody. The MET knockout cell lysate was generated from individual clone of MET knockout cell line (SKU. CL0018998301).
MET antibody showed specificity by CRISPR-mediated knockout in western blot. Western blot analysis of MET antibody was performed by loading each 10 µg of MET knockout cell lysate and wild-type HeLa cell lysate. The samples were separated by SDS-PAGE and immunoblotted with anti-MET monoclonal antibody. MET was detected at ~145 kDa. The result confirming specificity of the antibody. Data were normalized to a anti-ß-actin as a loading control(1:500).