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NS3-CMPK-hLBR1TM-mEGFP
Tissue
宫颈内口
Disease
腺癌
Metastatic site
原发肿瘤部位(宫颈管/宫颈)
Synonyms
HeLa R19 FlpIn TREx H2B-Cherry/NS3-CMPK-hLBR1TM-mEGFP

Documents

NS3-CMPK-hLBR1TM-mEGFP

Cell line · Product number 300986

This clonal stable cell line was generated by transfection of a circular plasmid and by Flp recombination followed by drug resistance selection.

US$800.00excl. tax and shipping
In stock
Lot
Recommended (freshest lot)
1

01General information

Description
这种克隆稳定细胞系是通过转染环形质粒和 Flp 重组,然后进行抗药性选择产生的。
Tissue
宫颈内口
Disease
腺癌
Metastatic site
原发肿瘤部位(宫颈管/宫颈)
Applications
核膜重塑;层蛋白B受体(LBR)生物学;多西环素诱导的基因表达;HeLa Kyoto FlpIn TREx系统;染色质与核膜的相互作用;活细胞成像;利用NS3介导的邻近标记进行的条件性耗竭研究
Synonyms
  • HeLa R19 FlpIn TREx H2B-Cherry/NS3-CMPK-hLBR1TM-mEGFP

02Classification

Organism
Human
Sex
Female
Growth properties
Adherent
GMO status
GMO-S1
GMO status details
GMO-S1: This HeLa R19 FlpIn TREx derivative contains Flp recombinase-integrated constructs encoding H2B-mCherry and doxycycline-inducible NS3-CMPK-hLBR1TM-mEGFP, with G418 resistance marker. This classification applies only within Germany and may differ elsewhere.
Biosafety level
1

03Characteristics

Morphology
上皮样
Cell type
上皮细胞

04Identifiers

Citation
NS3-CMPK-hLBR1TM-mEGFP(Cytion 目录号 300986)
NCBI TaxID
9606
Cellosaurus
CVCL_UR51

05Expression and susceptibility

Protein expression
H2B-mCherry 和 DOx 诱导型 NS3-CMPK-hLBR1TM-mEGFP

06Handling 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、0.5 mg/mL G418
Dissociation reagent
Accutase
Subculturing
去除粘附细胞上的旧培养基,用不含钙和镁的 PBS 冲洗。T25 烧瓶用 3-5 毫升 PBS,T75 烧瓶用 5-10 毫升。然后用 Accutase 完全覆盖细胞,T25 烧瓶用 1-2 毫升,T75 烧瓶用 2.5 毫升。让细胞在室温下孵育 8-10 分钟,使其分离。孵育后,用 10 毫升培养基轻轻混合细胞使其重悬,然后用 300xg 离心 3 分钟。弃去上清液,用新鲜培养基重悬细胞,并将其转移到已装有新鲜培养基的新烧瓶中。
Fluid renewal
每周 2 至 3 次
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 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

  7. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.

  8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

07Quality control and cryopreservation

Sterility

使用基于 PCR 的检测方法和基于发光的支原体检测方法排除支原体污染。

为确保没有细菌、真菌或酵母菌污染,每天都要对细胞培养物进行目视检查。

Freeze medium
我们使用完全生长培养基(包括 FBS)+10% DMSO 或 CM-1(Cytion 商品目录编号 800100)作为冷冻保存培养基,以获得足够的解冻后存活率,CM-1(Cytion 商品目录编号 800100)含有优化的渗透保护剂和代谢稳定剂,可提高恢复能力并减少冷冻引起的应激。
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。

08Donor

Donor age
30年
Ethnicity
非洲裔美国人