HK EGFP-H2B Cells

Categories: Reporter-labeled cells Description: 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. Organism: Human Tissue: Cervix Disease: Carcinoma Synonyms: HeLa Kyoto H2B-EGFP, HeLa Kyoto H2B EGFP, HeLa-H2B-GFP Age: 30 years Gender: Female Ethnicity: African American Morphology: Epithelial-like cells with mosaic stone shape Growth Properties: Monolayer, adherent Citation: HK EGFP-H2B (Cytion catalog number 300673) Biosafety Level: 1 Ncbi_ Taxid: 9606.0 Cellosaurus Accession: CVCL_1D63 Protein Expression: EGFP-H2B: Location/Gene: 1..589 / Pcmv, 613..1329 / EGFP, 1387..1764 / H2B, 3001..3795 / KanR/NeoR Products: CMV Promotor, Histone H2B, Neomycin, Phosphotransferase Culture Medium: 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) Supplements: Supplement the medium with 10% FBS Dissociation Reagent: Accutase Subculturing: 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. Seeding Density: 1 x 104 cells/cm2 Fluid Renewal: 2 to 3 times per week Post Thaw Recovery: 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. Freeze Medium: As a cryopreservation medium, 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. Thawing And Culturing Cells: 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. Sterility: 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. Safety Precautions: When planning to store a cryovial in liquid nitrogen for future thawing, it is mandatory to adhere to stringent safety measures. Appropriate protective gloves and clothing are essential, and the use of a face mask or safety goggles is required during the transfer of frozen samples to or from the liquid nitrogen tank. This is to mitigate the risk of injury from potential cryovial explosions upon removal, which can result in the projection of sharp fragments. Warranty: We stand by the promise of delivering products with high cell viability and robust culture performance. To achieve the best results, please make sure you follow the storage and culture instructions detailed in the product information sheet closely. Your adherence to these guidelines is key to success. Subject To Third- Party Agreements: Please note that this cell line is not available under a standard Cytion MTA, as it requires a third-party agreement and/or is subject to negotiation with the original licensor. Required Product 1: 820300a Required Product 3: 860015.0 Required Product 4: 830100.0