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HeLa EB3-GFP Kirc 14 09/27/24
Tissue
子宫颈
Disease
宫颈腺癌(HeLa背景);稳定转染了GFP-EB3(微管正端报告基因)

Documents

HeLa EB3-GFP Kirc 14 09/27/24

Cell line · Product number 305718

HeLa EB3-GFP Kirc 14 is a genetically modified derivative of the human HeLa cervical carcinoma cell line, engineered to stably express a GFP-tagged EB3 (e...

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01General information

Description

HeLa EB3-GFP Kirc 14 是人类 HeLa 宫颈癌细胞系的基因改造衍生物,经过工程改造可稳定表达一个带有 GFP 标签的 EB3(末端结合蛋白 3)融合构建体,其中 其中“Kirc 14”指从转染细胞池中筛选出的特定亚克隆(克隆14)。EB3是末端结合蛋白家族的成员,能够追踪生长中的微管正端,并作为活细胞中微管动态行为的报告蛋白。 GFP-EB3融合蛋白在聚合中的微管尖端产生特征性的彗星状荧光信号,从而能够对微管动力学、细胞内运输以及细胞骨架的组织结构进行实时活细胞成像。

HeLa EB3-GFP Kirc 14 用于细胞生物学研究,可对微管正端动力学进行活细胞成像,研究有丝分裂纺锤体组装和着丝点-微管相互作用,探究细胞内货物运输,并评估影响微管聚合动力学的化合物。 稳定的GFP-EB3亚克隆表达可在多次传代过程中保持一致的荧光强度和分布模式,因此第14号克隆适用于定量成像实验以及微管靶向药物的快速筛选。

Tissue
子宫颈
Disease
宫颈腺癌(HeLa背景);稳定转染了GFP-EB3(微管正端报告基因)
Applications
活细胞微管动力学成像;有丝分裂纺锤体研究;着丝粒-微管相互作用研究;细胞内运输;微管靶向药物筛选

02Classification

Organism
Human
Growth properties
Adherent
GMO status
GMO-S1
GMO status details
GMO-S1: This HeLa EB3 GFP KIRC14 line contains a GFP-tagged EB3 construct for microtubule plus-end tracking in live-cell imaging. 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小时

04Identifiers

Citation
HeLa EB3-GFP Kirc 14 2024年9月27日(Cytion 产品编号 305718)
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
Accutase,10分钟,37°C
Subculturing
从贴壁细胞中移除旧培养基,并用PBS洗涤。加入Accutase,室温孵育8-10分钟,用培养基重悬细胞,以300×g离心3分钟,然后转移至装有筛选培养基的新培养瓶中。
Split ratio
1 到 5
Seeding density
1 至 3 × 10⁴ 个细胞/cm²
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 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。