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CDKN2A Knockout 293T Cell Line, Homozygous (RM01875)

Genome sequence analysis of PCR products from parental (WT) and CDKN2A knockout (KO) 293T cells, using sanger sequencing.

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货号: RM01875
促销价:   ¥10000
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详细信息

靶点
CDKN2A
细胞系
293T
突变描述
CDKN2A Knockout 293T Cell Line is engineered from 293T cell line with Gene-Editing Technology.
Allele-1:41bp deletion in exon2
Allele-2:41bp 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
细胞数目
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 generates several transcript variants which differ in their first exons. At least three alternatively spliced variants encoding distinct proteins have been reported, two of which encode structurally related isoforms known to function as inhibitors of CDK4 kinase. The remaining transcript includes an alternate first exon located 20 Kb upstream of the remainder of the gene; this transcript contains an alternate open reading frame (ARF) that specifies a protein which is structurally unrelated to the products of the other variants. This ARF product functions as a stabilizer of the tumor suppressor protein p53 as it can interact with, and sequester, the E3 ubiquitin-protein ligase MDM2, a protein responsible for the degradation of p53. In spite of the structural and functional differences, the CDK inhibitor isoforms and the ARF product encoded by this gene, through the regulatory roles of CDK4 and p53 in cell cycle G1 progression, share a common functionality in cell cycle G1 control. This gene is frequently mutated or deleted in a wide variety of tumors, and is known to be an important tumor suppressor gene. [provided by RefSeq, Sep 2012]