DFFA Knockout 293T Cell Line, Homozygous (RM02243)

说明书收藏

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

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货号: RM02243

详细信息

靶点

DFFA

细胞系

293T

突变描述

DFFA Knockout cell line is engineered from 293T cell line with Gene-Editing Technology.
Allele-1:83bp deletion in exon2
Allele-2:83bp 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

Apoptosis is a cell death process that removes toxic and/or useless cells during mammalian development. The apoptotic process is accompanied by shrinkage and fragmentation of the cells and nuclei and degradation of the chromosomal DNA into nucleosomal units. DNA fragmentation factor (DFF) is a heterodimeric protein of 40-kD (DFFB) and 45-kD (DFFA) subunits. DFFA is the substrate for caspase-3 and triggers DNA fragmentation during apoptosis. DFF becomes activated when DFFA is cleaved by caspase-3. The cleaved fragments of DFFA dissociate from DFFB, the active component of DFF. DFFB has been found to trigger both DNA fragmentation and chromatin condensation during apoptosis. Two alternatively spliced transcript variants encoding distinct isoforms have been found for this gene. [provided by RefSeq, Jul 2008]

基因ID

1676

Swiss Prot

O00273

别名

DFF-45; DFF1; ICAD

研究领域