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HeLa EB3-GFP Kirc 14 09/27/24
Tissue
子宮頸
Disease
子宮頸腺癌(HeLa 背景);穩定轉染 GFP-EB3(微管正端報告基因)

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...

US$800.00excl. tax and shipping
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Lot
Recommended (freshest lot)
  • CoA
1

01General information

Description

HeLa EB3-GFP Kirc 14 是人類 HeLa 子宮頸癌細胞系的基因改造衍生物,經工程化設計可穩定表達帶有 GFP 標記的 EB3(末端結合蛋白 3)融合構建體,其中 其中「Kirc 14」代表從轉染細胞群中篩選出的特定亞克隆(第 14 號克隆)。EB3 屬於末端結合蛋白家族,能追蹤生長中微管的正端,並用作活細胞中微管動態行為的報告蛋白。 GFP-EB3 融合蛋白會在聚合中的微管頂端產生特有的彗星狀螢光訊號,從而能夠對微管動力學、細胞內運輸及細胞骨架組織進行即時活細胞成像。

HeLa EB3-GFP Kirc 14 廣泛應用於細胞生物學研究,用於微管正端動態的活細胞成像、有絲分裂紡錘體組裝及動點-微管相互作用的研究、細胞內貨物運輸的探究,以及評估影響微管聚合動態的化合物。 穩定的 GFP-EB3 亞克隆表達可在多次傳代過程中提供一致的螢光強度與分布模式,使第 14 號克隆適用於定量成像分析及微管靶向藥物的篩選。

Tissue
子宮頸
Disease
子宮頸腺癌(HeLa 背景);穩定轉染 GFP-EB3(微管正端報告基因)
Applications
活細胞微管動力學成像;有絲分裂紡錘體研究;動點-微管相互作用研究;細胞內運輸;微管靶向藥物篩選

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
上皮樣
Cell type
上皮細胞
Doubling time
約 24 小時

04Identifiers

Citation
HeLa EB3-GFP Kirc 14 2024年9月27日 (Cytion 型號 305718)
NCBI TaxID
9606

05Handling and culture

Culture medium
DMEM,含 4.5 g/L 葡萄糖、4 mM L-谷氨醯胺、3.7 g/L NaHCO₃ 及 1.0 mM 丙酮酸鈉(Cytion 產品編號 820300a)
Incubation atmosphere

37°C,5% CO₂,加濕環境。

Supplements
在培養基中加入 10% 的胎牛血清(FBS)
Dissociation reagent
Accutase,10 分鐘,37°C
Subculturing
將附著細胞中的舊培養基移除,並以PBS沖洗。加入Accutase覆蓋細胞,於室溫下孵育8至10分鐘,再以培養基重懸細胞,以300×g離心3分鐘,最後將細胞轉移至裝有篩選培養基的新培養瓶中。
Split ratio
1 至 5
Seeding density
1 至 3 × 10⁴ 個細胞/cm²
Fluid renewal
每週 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
作為冷凍保存培養基,我們使用完全生長培養基加上 10% 的二甲基亞砜(DMSO),以確保解凍後的細胞存活率達到適當水準。
Storage conditions

若需長期保存,請將試管置於溫度約為 −150 至 −196 °C 的氣相液氮中。僅在轉移至液氮之前,可將試管暫時存放於 −80 °C 環境中,作為短期過渡步驟。

Shipping conditions

冷凍保存的細胞系會以乾冰冷藏,並裝於經驗證的隔熱包裝中,包裝內含有足夠的冷媒,以確保在運輸過程中溫度維持在約 −78 °C。收到貨物後,請立即檢查容器,並盡快將試管轉移至適當的儲存場所。