SKP2 Knockout HCT116 Cell Line, Homozygous

Cell Lines & Derivatives

This gene encodes a member of the F-box protein family which is characterized by an approximately 40 amino acid motif, the F-box. The F-box proteins constitute one of the four subunits of ubiquitin protein ligase complex called SCFs (SKP1-cullin-F-box), which function in phosphorylation-dependent ubiquitination. The F-box proteins are divided into 3 classes: Fbws containing WD-40 domains, Fbls containing leucine-rich repeats, and Fbxs containing either different protein-protein interaction modules or no recognizable motifs. The protein encoded by this gene belongs to the Fbls class; in addition to an F-box, this protein contains 10 tandem leucine-rich repeats. This protein is an essential element of the cyclin A-CDK2 S-phase kinase. It specifically recognizes phosphorylated cyclin-dependent kinase inhibitor 1B (CDKN1B, also referred to as p27 or KIP1) predominantly in S phase and interacts with S-phase kinase-associated protein 1 (SKP1 or p19). In addition, this gene is established as a protooncogene causally involved in the pathogenesis of lymphomas. Alternative splicing of this gene generates three transcript variants encoding different isoforms. [provided by RefSeq, Jul 2011]

1 vial parental cell line and 1 vial knockout cell line

Upon arrival, it should be maintained in DMEM medium with 10%(v/v) fetal bovine serum and 100U penicillin-streptomycin, at 37°C with 5% CO₂ condition.
1. Thaw the vial in 37°C water bath , and shake it to melt as soon as possible.
2. Transfer the cell suspension to a 15mL conical tube with pre-warmed 5mL complete me-
        dium and centrifuge 1000rpm for approximately 5 minutes at room temperature.
3. Remove and discard the supernatant.
4. Resuspend the cell pellet with 1mL pre-warmed complete medium and seed in 10cm dish.
5. Add 8-10mL of complete medium.
6. Incubate the culture at 37°C incubator with 5% CO₂.
7. A subcultivation ratio of 1:2-1:4 is recommended.