Skip to content
HK EGFP-LaminA/H2B-mCherry
Tissue
子宮頸
Disease
癌
Synonyms
HeLa Kyoto EGFP-LaminA and H2B-mCherry

Documents

HK EGFP-LaminA/H2B-mCherry

Cell line · Product number 300921

The HK EGFP-LaminA/H2B-mCherry cell line is a genetically engineered Hela Kyoto derived cell model developed to facilitate advanced studies in nuclear dyn...

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

01General information

Description

HK EGFP-LaminA/H2B-mCherry 細胞系是一種源自 Hela Kyoto 的基因工程細胞模型,旨在促進對活細胞內核動力學及染色質組織結構的深入研究。 該細胞系表達兩種融合蛋白:EGFP(增強型綠色螢光蛋白)與 Lamin A 的融合蛋白,以及 mCherry(一種紅色螢光蛋白)與組蛋白 H2B 的融合蛋白。 EGFP-Lamin A 融合蛋白可突顯核膜,並能使細胞週期進展過程中或各種實驗條件下核結構的變化得以視覺化。與此同時,H2B-mCherry 融合蛋白會與 DNA 結合,並發出鮮豔的紅色螢光以標記染色質,從而能夠即時觀察有絲分裂和間期期間的染色體過程。

這些細胞對於即時成像應用具有無可替代的價值,包括核完整性、DNA 複製及細胞老化等研究,以及針對核結構受損疾病(如癌症和核膜蛋白病)的研究。 該細胞系具備雙色螢光特性,可同時顯影核膜與染色質,有助於全面理解核-細胞質相互作用以及染色質的時空組織。 這些特性使其成為分子生物學研究與細胞生物物理學的重要工具,有助於深入了解基因表達調控、核組織結構及細胞週期的機制。

Tissue
子宮頸
Disease
癌
Synonyms
  • HeLa Kyoto EGFP-LaminA and H2B-mCherry

02Classification

Organism
Human
Sex
Female
Growth properties
Adherent
GMO status
GMO-S1
GMO status details
GMO-S1: This HeLa Kyoto line contains EGFP-Lamin A and H2B-mCherry constructs enabling dual-color imaging of nuclear lamina and chromatin. This classification applies only within Germany and may differ elsewhere.
Biosafety level
1

03Characteristics

Morphology
呈馬賽克狀石塊形狀的類上皮細胞

04Identifiers

Citation
HK EGFP-LaminA/H2B-mCherry(Cytion 型號 300921)
NCBI TaxID
9606
Cellosaurus
CVCL_1D62
Depositor
艾倫伯格實驗室(EMBL)

05Expression and susceptibility

Protein expression
EGFP-LaminA/H2B-mCherry
Products
組蛋白 H2B

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% 的胎牛血清(FBS)
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。
Seeding density
1 × 10⁴ 個細胞/cm²
Fluid renewal
每週 2 至 3 次
Post-thaw recovery
解凍後,將細胞以 5 × 10⁴ 個細胞/cm² 的密度接種於培養皿中,並讓細胞從冷凍過程中恢復,並附著至少 24 小時。
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