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- 博尔森生物
- BES2097K-1-4PL
- 中国/上海
- 2025年11月26日
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- 详细信息
- 文献和实验
- 技术资料
- 品系：
人
- 细胞类型：
正常细胞
- 肿瘤类型：
否
- 供应商：
上海博尔森生物科技有限公司
- 库存：
大量
- 英文名：
Cytochrome C Knockout Hela Cell Ripa Lysate (Pool)
- 生长状态：
详见操作说明
- 年限：
12个月
- 运输方式：
4℃
- 器官来源：
人
- 是否是肿瘤细胞：
否
- 细胞形态：
人
- 免疫类型：
详见操作说明
- 物种来源：
人
- 相关疾病：
详见操作说明
- 组织来源：
详见操作说明
- 规格：
50μL, 2μg/μl
Cytochrome C Knockout Hela Cell Ripa Lysate (Pool)
Gene Symbol CYCS
Synonyms CytC; CYC; CYCS; Cytochrome c somatic; HCS; CYC_HUMAN; Cytochrome-c; MSA06; THC4.
gRNA Species Human
Parental Cell line Hela
Knockout Region E2
Knockout Validation PCR + Sanger Sequencing
Mutation Description Knockout achieved by using CRISPR/Cas9, Deletion of the selection cassette in exon 2.
Applications: WB
Packaging 1 vial knockout cell Lysate and 1 vial parental cell Lysate.
Shipping Conditions 4℃
Storage Store at -20°C for 6 months storage or -80°C for long-term storage.
Notes This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.
Background Cytochrome C is an electron transporting protein that resides within the intermembrane space of the mitochondria, where it plays a critical role in the process of oxidative phosphorylation and production of cellular ATP. An increasing amount of interest has been directed toward the role which cytocrome C has been demonstrated to play in apoptotic processes. Following exposure to apoptotic
stimuli, cytochrome C is rapidly released from the mitochondria into the cytosol, an event which may be required for the completion of apoptosis in some systems. Cytosolic cytochrome C functions in the activation of caspase 3, an ICE family molecule that is a key effector of apoptosis.
Protocol Lysates are made using Ripa buffer with 1mM Phenylmethylsulfonyl fluoride (PMSF).
Proteins in this buffer are denatured.To use as WB control, SDS-PAGE loading buffer should be added to the lysate and boiled for 5 minutes, then loaded with gel.
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文献和实验多篇 SCI 发现用 RIPA 提蛋白存在问题，我们该如何应对？
7 [3]（下图），表明用 RIPA 裂解液提取这些特定蛋白质的效率十分低。近年来，越来越多的研究者密切关注 RIPA 不可溶组分中的蛋白质组分，以及它们对下游实验和整体数据的影响。Bai 和 Laiho 用 RIPA 裂解液从 Hela 细胞核中提取蛋白，发现可溶性组分和不可溶性组。分的蛋白质图谱差异很大，表明蛋白质的丢失不成正比 [4] （下图）。Mukhopadhyay 等 [5] 从突变小鼠的乳腺上皮细胞中提取总蛋白，发现在 RIPA 不可溶组分中，通过 WB 很容易检测到 EGFR, HSP
Gene Knockout Techniques(1) The Knockout Process
). Spin cell debris to bottom of wells (2000 RPM/2 minutes). Pool by sampling 75 ul from each well of a plate with the Hydra and dispensing 70 ul into a sterile 8-channel pooling block (this looks like a tiny little swimming pool with 8 horizontal lanes
that all the cells are detached. Prolong incubation if needed. Add into each dish 8 ml of pre-warmed medium, pipette up and down so that all the cells are collected as a single-cell suspension. Pool all the harvested cells into 2x 50ml Falcon tubes. For some strains
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