HK Mad2-LAP/H2B-mCherry Cells

USD 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).

Los precios no incluyen impuestos ni gastos de envío

cryovial

Número de producto: 300920

Ficha de datos de seguridad

Ficha del producto

Certificado de análisis (CoA)

Descripción	The HK Mad2-LAP/H2B-mCherry cell line is a genetically engineered cell model extensively utilized for studying chromosome segregation and the spindle assembly checkpoint during mitosis. These cells are derived from HeLa Kyoto cells, a robust human cell line originally taken from a cervical carcinoma. The HK Mad2-LAP (LAP-tagged Mad2) aspect of the cell line facilitates the visualization and functional analysis of the Mad2 protein, a critical component of the spindle assembly checkpoint that prevents anaphase onset until all chromosomes are properly aligned at the metaphase plate. Incorporation of H2B-mCherry, where histone H2B is tagged with the mCherry fluorescent protein, allows for real-time imaging of chromatin dynamics during cell division. This feature makes the HK Mad2-LAP/H2B-mCherry cell line an excellent tool for high-resolution live-cell imaging techniques to observe chromosomal movements and mitotic progression in human cells under various experimental conditions. The use of fluorescent tags aids in precise tracking and quantification, thereby providing valuable insights into the molecular mechanisms governing cell cycle regulation and chromosomal stability.
Organismo	Human
Tejido	Cervix
Enfermedad	Carcinoma
Sinónimos	HeLa Kyoto Mad2-LAP and H2B-mCherry, HeLa Kyoto Mad2-LAP

Edad	30 years
Género	Female
Origen étnico	African American
Morfología	Epithelial-like cells with mosaic stone shape
Propiedades de crecimiento	Monolayer, adherent

Referencia	HK Mad2-LAP/H2B-mCherry (Cytion catalog number 300920)
Nivel de bioseguridad	1
NCBI_TaxID	9606
N.º de acceso de Cellosaurus	CVCL_1D65
Depositante	The Ellenberg Lab (EMBL)
Estado de los OGM	GMO-S1: This HeLa Kyoto line contains Mad2-LAP and H2B-mCherry constructs enabling visualization of spindle checkpoint dynamics. This classification applies only within Germany and may differ elsewhere.

Expresión de proteínas	Mad2-LAP/H2B-mCherry

Medio de cultivo	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
Reactivo de disociación	Accutase
Subcultivo	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.
Densidad de siembra	1 x 10⁴ cells/cm²
Renovación de fluidos	2 to 3 times per week
Recuperación tras el deshielo	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.
Medio de congelación	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.
Descongelación y cultivo 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.
Atmósfera de incubación	37°C, 5% CO₂, humidified atmosphere.
Condiciones de envío	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.
Condiciones de almacenamiento	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.

Esterilidad	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.