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NS3-CMPK-hLBR1TM-mEGFP
Tissue
子宮頸內口
Disease
腺癌
Metastatic site
原發性腫瘤部位(子宮頸內口/子宮頸)
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
此克隆穩定細胞系是透過轉染環狀質體,經由 Flp 重組,並隨後進行抗藥性篩選而建立的。
Tissue
子宮頸內口
Disease
腺癌
Metastatic site
原發性腫瘤部位(子宮頸內口/子宮頸)
Applications
核膜重塑;層蛋白 B 受體(LBR)生物學;多西環素誘導性基因表達;HeLa Kyoto FlpIn TREx 系統;染色質與核膜的相互作用;活細胞成像;利用 NS3 介導的鄰近標記進行的條件性耗竭研究
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
上皮樣
Cell type
上皮細胞

04Identifiers

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

05Expression and susceptibility

Protein expression
H2B-mCherry 及 DOx 誘導型 NS3-CMPK-hLBR1TM-mEGFP

06Handling 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% FBS 及 0.5 mg/mL G418
Dissociation reagent
Accutase
Subculturing
從貼壁細胞中移除舊培養基,並用不含鈣和鎂的PBS洗滌細胞。T25培養瓶使用3-5毫升PBS,T75培養瓶則使用5-10毫升。 接著,用 Accutase 完全覆蓋細胞,T25 培養瓶使用 1–2 毫升,T75 培養瓶則使用 2.5 毫升。將細胞於室溫下孵育 8–10 分鐘,使其脫附。 孵育後,將細胞與 10 毫升培養基輕柔混合以使其重新懸浮,然後以 300xg 離心 3 分鐘。棄去上清液,將細胞重新懸浮於新鮮培養基中,並轉移至已預先加入新鮮培養基的新培養瓶中。
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 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

透過基於 PCR 的檢測及基於發光原理的支原體檢測方法,排除支原體污染的可能性。

為確保無細菌、真菌或酵母菌污染,每日均會對細胞培養物進行目視檢查。

Freeze medium
作為冷凍保存培養基,我們使用完全生長培養基(含 FBS)+ 10% DMSO 以確保解凍後的存活率,或使用 CM-1 (Cytion 型號 800100),該培養基含有經優化的滲透保護劑和代謝穩定劑,可提升細胞恢復率並減輕冷凍誘導的壓力。
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。

08Donor

Donor age
30年
Ethnicity
非裔美國人