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Stable HEK293 Cells-Rpn11-HTBH
Tissue
肾脏
Disease
转基因/永生化胎儿肾脏(HEK293背景;Rpn11-HTBH转基因)

Documents

Stable HEK293 Cells-Rpn11-HTBH

Cell line · Product number 305719

Stable HEK293 Cells-Rpn11-HTBH is a stably transfected derivative of the HEK293 (Human Embryonic Kidney 293) cell line, engineered to express a tagged ver...

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Lot
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01General information

Description

稳定HEK293-Rpn11-HTBH细胞是HEK293 (人胚胎肾293)细胞系的稳定转染衍生物,经工程改造可表达带标签的Rpn11(又称PSMD14或POH1),该蛋白是26S蛋白酶体盖子复合体的去泛素化酶亚基。 Rpn11是一种Zn²⁺依赖性的JAMM结构域去泛素化酶,可在蛋白酶体降解过程中从与蛋白酶体结合的底物上去除泛素链。 HTBH标签(六组氨酸-TEV-生物素受体肽-六组氨酸)可在天然条件下实现含Rpn11复合物的亲和纯化,因此该细胞系特别适用于蛋白酶体复合物的纯化及相互作用组研究。

该细胞系适用于 26S 蛋白酶体生物学、泛素-蛋白酶体通路(UPS)调控、Rpn11/PSMD14 在蛋白质质量控制中的功能、蛋白酶体组装与动力学以及蛋白酶体抑制剂作用机制的研究。 它还用于天然蛋白酶体复合物的亲和纯化,并作为研究蛋白酶体背景下去泛素酶生物学的模型。HTBH 标签系统可通过基于链霉亲和素的拉下法,对生物素标记的复合物进行高度严格的纯化。

Tissue
肾脏
Disease
转基因/永生化胎儿肾脏(HEK293背景;Rpn11-HTBH转基因)
Applications
26S蛋白酶体的生物学;Rpn11/PSMD14的功能;泛素-蛋白酶体通路;蛋白酶体复合物的纯化;去泛素酶的生物学;HTBH标签亲和纯化;蛋白酶体相互作用组研究

02Classification

Organism
Human
Growth properties
Adherent
GMO status
GMO-S1
GMO status details
GMO-S1: This HEK293 derivative contains a stably integrated Rpn11-HTBH expression cassette (hexahistidine-TEV-biotin acceptor peptide-hexahistidine tagged Rpn11/PSMD14). This classification applies only within Germany and may differ elsewhere.
Biosafety level
1

03Characteristics

Morphology
上皮样
Cell type
上皮细胞
Doubling time
约24至36小时

04Identifiers

Citation
稳定HEK293细胞-Rpn11-HTBH(Cytion产品编号 305719)
NCBI TaxID
9606

05Handling and culture

Culture medium
DMEM,w:4.5 克/升葡萄糖,w:4 毫摩尔 L-谷氨酰胺,w:3.7 克/升 NaHCO3,w:1.0 毫摩尔丙酮酸钠(Cytion 编号 820300a)。
Incubation atmosphere

37°C, 5%CO2, 加湿环境。

Supplements
在培养基中添加 10% FBS
Dissociation reagent
大多数细胞在PBS中即可脱落,如有必要,可在室温下加入Accutase处理5分钟
Subculturing
移除培养基,用不含钙和镁的PBS洗涤,加入Accutase覆盖细胞,在室温下孵育8–10分钟,用培养基重悬细胞,以300×g离心3分钟,弃去上清液,将细胞接种到新鲜培养基中。
Split ratio
1 到 10
Seeding density
2 至 4 × 10⁴ 个细胞/cm²
Fluid renewal
每 2 至 3 天
Thawing and culturing
  1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

  2. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.

  3. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.

  4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

  5. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.

  6. Centrifuge the mixture at 200 x g for 5 minutes, carefully discard the supernatant containing freezing medium.

  7. Follow the procedure described under Post-Thaw Recovery

06Quality control and cryopreservation

Freeze medium
作为冷冻保存培养基,我们使用完全生长培养基+10% DMSO,以获得足够的解冻后存活率。
Storage conditions

如需长期保存,可将样品瓶放入气相液氮中,温度约为 -150 至 -196 ℃。在-80 °C下保存只能作为转入液氮前的短暂过渡。

Shipping conditions

冷冻保存的细胞系用干冰装运,并用经过验证的隔热包装和足够的制冷剂在整个运输过程中保持约 -78 °C 的温度。收到货后,应立即检查容器,并立即将小瓶转移到适当的储藏室。