HK EB3-EGFP Cells

USUSD$800.00*

Products are shipped frozen on dry ice in cryotubes. Each cryotube typically contains 3 × 10⁶ cells for adherent lines or 5 × 10⁶ cells for suspension lines (refer to the batch CoA for details).

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cryovial

產品編號： 300668

安全資料表

產品說明書

說明	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 Green Fluorescent Protein (EGFP). This cell line is commonly utilized in research focused on understanding microtubule dynamics due to the fluorescent tagging of EB3, a protein that associates with the plus ends of microtubules. The expression of EGFP provides a fluorescent marker that allows for real-time visualization of microtubule behavior in live cells under a fluorescence microscope. This cell line is particularly valuable in cell biology and cancer research, where understanding the mechanics of cell division and intracellular transport is crucial. The stable expression of EB3-EGFP does not interfere with the normal functions of the microtubules, making these cells a reliable tool for detailed studies of cellular processes that depend on microtubule dynamics.
生物體	Human
組織	Cervix
疾病	Carcinoma
同義詞	HeLa Kyoto EB3-EGFP, HeLa Kyoto EB3 EGFP, HeLa Kyoto EGFP-EB3

年齡	30 years
性別	Female
族裔	African American
形態學	Epithelial-like cells with mosaic stone shape
生長特性	Monolayer, adherent

引用	HK EB3-EGFP (Cytion catalog number 300668)
生物安全等級	1
NCBI_TaxID	9606
Cellosaurus 編號	CVCL_1D61
存款人	The Ellenberg Lab (EMBL)
轉基因狀態	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.

蛋白質表達	MEGFP (microtubule End-binding protein 3 mEGFP tagged): Location/Gene: 1..589 / Pcmv, 652..1497 / EB3, 1516..2235 / EGFP, 3466..4260 / KanR/NeoR
產品	CMV Promotor EB3, Neomycin, Phosphotransferase

培養基	DMEM, w: 4.5 g/L Glucose, w: 4 mM L-Glutamine, w: 3.7 g/L NaHCO3, w: 1.0 mM Sodium pyruvate (Cytion article number 820300a)
營養補充品	Supplement the medium with 10% FBS
解離試劑	Accutase
傳代培養	Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium.
播種密度	1 x 10⁴ cells/cm²
流體更新	2 to 3 times per week
解凍後的恢復	After thawing, plate the cells at 5 x 10⁴ cells/cm² and allow the cells to recover from the freezing process and to adhere for at least 24 hours.
冷凍培養基	As a cryopreservation medium, we use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress.
細胞解凍與培養	Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit. 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. 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. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening. 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. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium. 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. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.
培養環境	37°C, 5% CO₂, humidified atmosphere.
運送條款	Cryopreserved cell lines are shipped on dry ice in validated, insulated packaging with sufficient refrigerant to maintain approximately −78 °C throughout transit. On receipt, inspect the container immediately and transfer vials without delay to appropriate storage.
儲存條件	For long-term preservation, place vials in vapor-phase liquid nitrogen at about −150 to −196 °C. Storage at −80 °C is acceptable only as a short interim step before transfer to liquid nitrogen.

不孕症	Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods. To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections.

HK EB3-EGFP Cells

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