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HK EB3-EGFP
Tissue
子宫颈
Disease
癌症
Synonyms
HeLa Kyoto EB3-EGFP, HeLa Kyoto EB3 EGFP, HeLa Kyoto EGFP-EB3

Documents

HK EB3-EGFP

Cell line · Product number 300668

Hela Kyoto EB3-EGFP is a derivative of the HeLa Kyoto cell line, specifically engineered to express the End-Binding Protein 3 (EB3) tagged with Enhanced G...

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01General information

Description

Hela Kyoto EB3-EGFP是HeLa Kyoto细胞系的一种衍生物,专门用于表达末端结合蛋白3(EB3)和增强型绿色荧光蛋白(EGFP)。EB3 是一种与微管正端结合的蛋白质,由于它的荧光标记,这种细胞系通常用于了解微管动力学的研究。EGFP 的表达提供了一种荧光标记,可在荧光显微镜下实时观察活细胞中微管的行为。

这种细胞系在细胞生物学和癌症研究中尤为重要,因为了解细胞分裂和细胞内运输的机理对研究至关重要。EB3-EGFP 的稳定表达不会干扰微管的正常功能,使这些细胞成为详细研究依赖于微管动力学的细胞过程的可靠工具。

Tissue
子宫颈
Disease
癌症
Synonyms
  • HeLa Kyoto EB3-EGFP
  • HeLa Kyoto EB3 EGFP
  • HeLa Kyoto EGFP-EB3

02Classification

Organism
Human
Sex
Female
Growth properties
Adherent
GMO status
GMO-S1
GMO status details
GMO-S1: This HeLa Kyoto EB3-EGFP line contains an EGFP-tagged EB3 construct for dynamic microtubule visualization. This classification applies only within Germany and may differ elsewhere.
Biosafety level
1

03Characteristics

Morphology
具有镶嵌石形状的上皮样细胞

04Identifiers

Citation
HK EB3-EGFP(Cytion 目录号 300668)
NCBI TaxID
9606
Cellosaurus
CVCL_1D61
Depositor
艾伦伯格实验室(EMBL)

05Expression and susceptibility

Protein expression
MEGFP(微管末端结合蛋白 3 mEGFP 标记):位置/基因:1...589 / Pcmv, 652...1497 / EB3, 1516...2235 / EGFP, 3466...4260 / KanR/NeoR
Products
CMV 促进因子 EB3、新霉素、磷基转移酶

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
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。
Seeding density
1 × 10⁴ 个细胞/平方厘米
Fluid renewal
每周 2 至 3 次
Post-thaw recovery
解冻后,将细胞以5×10⁴个细胞/平方厘米的密度接种于培养皿中,使细胞从冷冻过程中恢复并至少贴壁24小时。
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