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NRK-IBB-DiHcRed1
Tissue
腎臟
Synonyms
NRK IBB-DiHcRed1

Documents

NRK-IBB-DiHcRed1

Cell line · Product number 500671

NRK-IBB-DiHcRed1 is a modified cell line derived from normal rat kidney (NRK) cells, engineered to express the red fluorescent protein DiHcRed1. This modi...

US$800.00excl. tax and shipping
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1

01General information

Description

NRK-IBB-DiHcRed1 是一種源自正常大鼠腎臟(NRK)細胞的修飾細胞系,經基因工程改造後可表達紅色螢光蛋白 DiHcRed1。此項修飾使研究人員能夠利用螢光顯微鏡,即時追蹤並觀察細胞過程。 其穩定的紅色螢光非常適合活細胞成像,有助於進行細胞遷移、分裂及形態學的研究。

該細胞系保留了 NRK 細胞的典型特徵,包括上皮樣形態和正常的增殖能力,使其成為研究哺乳動物細胞行為的可靠模型。其紅色螢光還可與其他標記物進行多重檢測,從而增強其在細胞生物學、癌症研究及藥物篩選中的應用價值。

Tissue
腎臟
Synonyms
  • NRK IBB-DiHcRed1

02Classification

Organism
Rat
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
呈紡錘形的成纖維細胞樣細胞

04Identifiers

Citation
NRK-IBB-DiHcRed1(Cytion 型號 500671)
NCBI TaxID
10116
Cellosaurus
CVCL_AV95
Depositor
艾倫伯格實驗室(EMBL)

05Expression and susceptibility

Receptors expressed
表皮生長因子(EGF)、增殖刺激活性(MSA)
Protein expression
IBB-DiHcRed1:位置/基因:1..589 / Pcmv,656..916 / IBB,932..1615,1670..2356 / HcRed1,3587..4381 / KanR/NeoR
Products
CMV 啟動子 IBB(Ribbeck & Gorlich 2002)、新黴素、磷酸轉移酶、表皮生長因子、增殖刺激活性

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
倒掉舊培養基,並用PBS洗滌細胞。 加入新鮮配製並加熱至 37 攝氏度的 0.025% 胰蛋白酶/0.02% EDTA 溶液,並等待細胞脫落,此過程通常約需 5 分鐘。加入新鮮培養基以中和胰蛋白酶,然後將細胞混合物轉移至離心管中並進行離心。 離心後,倒去上清液,將細胞沉澱用新鮮培養基重懸,並將懸浮液轉移至新的培養瓶中。在培養基中加入 G418,使其最終濃度達到 0.5 mg/ml
Seeding density
2 至 4 × 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 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

Breed / subspecies
奧斯本·門德爾