Negative control for Western blotting application
What is KO cell lysate?
KO cell lysates are the cell homogenate in RIPA buffer made with double-knockout cell lines. The KO cell lines development was described in detail below. 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.
How was the KO cell lines developed?
OriGene teams up with EdiGene, a CRISPR innovator, who produces double-knockout genotype in a high-throughput manner. Unlike HAP1 cell line from Horizon, EdiGene KO cell lines are from the commonly used cell lines, such as HEK293T or HeLa, making the product more relevant for researchers.
What are the applications of the KO cell lysate?
The immediate utility is to serve as negative controls for western blots and ultimately to validate the specificity of antibodies.
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)
How is the Knockout cell line validated?
Each KO cell line carries mutation at both alleles of the target locus.
The validation was conducted at genomic level via PCR and sequencing using PCR primers strategically designed around the target site (P1 and P2), or on the inserted fragment (P3).
For example, for MET-Knockout cell (HeLa), one allele carries a 41bp deletion in exon 1, the other allele carries an 1.1 kb insertion of selection cassette with 3 X STOP for every frame. (Click to view the sample datasheet)