Antibody specificity has been a well-recognized problem in the research community. It causes unreliable or misinterpreted data and frustrates the research community. OriGene is a pioneer in creating a class of highly validated antibodies. Since 2013, OriGene has created UltraMABs, a growing list of monoclonal antibodies whose mono-reactivity are verified using a high-density protein chip with 17,000 overexpression lysates.
Starting 2017, OriGene approached the problem using a different strategy, using CRISPR to create a long list of Knockout Lysates. Cell lines with both alleles of a specific genes knocked out by CRISPR are called Knockout Cells, or KO Cells. Cell homogenates from the KO cells are the KO cell lysates. All of KO cell lines are validated at the genetic level using PCR, followed by sequencing. The 2 alleles are either interrupted by in-del or by insertion of the puro-resistant cassette.
The validation data is presented for each KO lysate product. Each KO cell lysate are supplied as a kit with 2 vials of lyophilized material: one for the KO cell lysate and the other for the parental Cell lysate, serving as the perfect control. Using these KO lysate, parent lysate pairs, OriGene has worked through our own monoclonal antibodies.Briefly the KO cell and its parental lysate were analyzed by western blotting. To qualify for a KO validated antibody, the target protein should be detected in the parental lane, but be absent in the knockout lane.
The same blot was stripped and re-probed with a loading control antibody, demonstrating equal loading of the 2 lysates. So far, over 100 of OriGene’s TrueMABs has been validated using this rigorous process. And the list is growing. They are prominently labeled by the KO validated logo. The KO-validated antibody list includes some popular names, such as HER2, IDH1, NOTCH1, c-MET, KI67, and many more. Both the KO cell lysate and the KO validated antibodies are searchable by typing a gene symbol into the search box.
Are you concerned with the validity of your antibody experiments? Come to OriGene to select the pre-validated antibodies or choose the KO lysates to validate your own antibodies. For more details, please visit the OriGene website.