HK EGFP-H2B Cells

US$ 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

Número do produto: 300673

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Descrição	The HK EGFP-H2B cell line is a genetically modified Hela Kyoto cell line used primarily for the study of chromatin dynamics and nuclear processes. This cell line expresses a fusion protein consisting of Enhanced Green Fluorescent Protein (EGFP) and histone H2B. The integration of EGFP into the H2B protein allows for the real-time visualization of chromatin in living cells under fluorescence microscopy, providing valuable insights into the spatial and temporal organization of the nucleus. The EGFP-H2B fusion facilitates numerous applications in cell biology, including the study of cell cycle progression, mitosis, and gene expression regulation. By observing the fluorescence patterns, researchers can identify and analyze phases of the cell cycle, chromosomal segregation, and structural changes within the nucleus. This cell line is derived from adult human cells, ensuring relevance to human biology, and is utilized in both basic biological research and more applied pharmaceutical studies. Additionally, the HK EGFP-H2B cell line serves as a crucial tool in epigenetics research. The ability to directly observe histone behaviors helps in understanding the epigenetic mechanisms that underlie gene expression and silencing, as well as the effects of various epigenetic modifiers. The cell line's robust application in live-cell imaging experiments makes it indispensable for detailed studies requiring dynamic cellular analysis.
Organismo	Human
Tecido	Cervix
Doença	Carcinoma
Sinônimos	HeLa Kyoto H2B-EGFP, HeLa Kyoto H2B EGFP, HeLa-H2B-GFP

Idade	30 years
Gênero	Female
Etnia	African American
Morfologia	Epithelial-like cells with mosaic stone shape
Propriedades de crescimento	Monolayer, adherent

Referência	HK EGFP-H2B (Cytion catalog number 300673)
Nível de biossegurança	1
NCBI_TaxID	9606
Número de acesso do Cellosaurus	CVCL_1D63
Depositante	The Ellenberg Lab (EMBL)
Situação em relação aos OGMs	GMO-S1: This HeLa Kyoto line contains an EGFP-H2B construct allowing real-time visualization of chromatin organization. This classification applies only within Germany and may differ elsewhere.

Expressão de proteínas	EGFP-H2B: Location/Gene: 1..589 / Pcmv, 613..1329 / EGFP, 1387..1764 / H2B, 3001..3795 / KanR/NeoR
Produtos	CMV Promotor, Histone H2B, Neomycin, Phosphotransferase

Meio de cultura	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)
Suplementos	Supplement the medium with 10% FBS
Reagente de dissociação	Accutase
Subcultura	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.
Densidade de semeadura	1 x 10⁴ cells/cm²
Renovação de fluidos	2 to 3 times per week
Recuperação pós-degelo	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.
Meio de congelamento	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.
Descongelamento e cultura de células	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.
Atmosfera de incubação	37°C, 5% CO₂, humidified atmosphere.
Condições de envio	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.
Condições de armazenamento	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.

Esterilidade	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 EGFP-H2B Cells

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Controle de Qualidade e Análise Molecular