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SKP2 Knockout HCT116 cell line, Homozygous (RM50184)

Genome sequence analysis of PCR products from parental (WT) and SKP2 knockout (KO) HCT116 cells, using sanger sequencing.

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货号: RM50184
促销价:   ¥10000
货    期:现货产品
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详细信息

靶点
SKP2
细胞系
HCT116
突变描述
SKP2 Knockout cell line is engineered from HCT116 cell line with Gene-Editing Technology.
Allele-1:137bp deletion in exon2
Allele-2:137bp deletion in exon2

Mammalian cells such as human, rat and mouse cells are normally diploid with two alleles.
Homozygote: both alleles were knocked out, mRNA has no signal, no expression of proteins.
Heterozygote: only one allele was knocked out, the mRNA transcript levels was decreased compared to wild type, and the protein expression levels was also lower than that of the wild type.
敲除验证
Sanger Sequencing
产品组成
1 vial parental cell line and 1 vial knockout cell line
裂解液量
1~5x106 cells/vial.
使用方法
Upon arrival, it should be maintained in DMEM medium with 10%(v/v) fetal bovine serum and 100U penicillin-streptomycin, at 37℃ with 5% CO2 condition.
1. Thaw the vial in 37℃ 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℃ incubator with 5% CO2.
7. A subcultivation ratio of 1:2-1:4 is recommended.
物种
Human
保存条件
Stored in liquid nitrogen for a long time less than -130℃. Minimizing freeze-thaw cycles.
运输条件
Dry ice

背景信息

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.

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