fr en es de it pt nl

Reactivos e instrumentos para inmunología, biología celular y biología molecular.

HK-2xZFN-Smc4-mEGFP Clone 82-68

Imprimir

Referencia 301576

embalaje : 1cryovial

Contact local distributor :

Teléfono : +1 850 650 7790

Marca : CLS Cell Lines Service

Contact local distributor :

Teléfono : +1 850 650 7790

HK-2xZFN-Smc4-mEGFP Clone 82-68 Cells

HK-2xZFN-Smc4-mEGFP Clone 82-68 Cells

HK-2xZFN-Smc4-mEGFP Clone 82-68 Cells

HK-2xZFN-Smc4-mEGFP Clone 82-68 Cells

HK-2xZFN-Smc4-mEGFP Clone 82-68 Cells

Description	The HK-2xZFN-Smc4-mEGFP Clone 82-68 cell line is a genetically engineered derivative of the human kidney proximal tubule epithelial cell line, HK-2. This cell line has been modified using zinc-finger nuclease (ZFN) technology to incorporate a fluorescently tagged version of the structural maintenance of chromosomes 4 (Smc4) protein, fused to the monomeric enhanced green fluorescent protein (mEGFP). The Smc4 protein is a critical component of the condensin complex, which plays a crucial role in chromosome condensation and segregation during cell division. The addition of mEGFP allows for real-time visualization and tracking of Smc4 dynamics within live cells, facilitating studies on chromosome architecture and the mechanisms governing mitosis. The HK-2xZFN-Smc4-mEGFP Clone 82-68 cell line is particularly valuable for research focused on cell cycle regulation, chromosomal behavior, and the structural maintenance of chromosomes. The stable expression of the mEGFP-tagged Smc4 protein ensures consistent fluorescence, making this cell line an excellent tool for live-cell imaging and quantitative analysis of condensin complex functions. Furthermore, the use of proximal tubule epithelial cells as the parental line provides a relevant model for kidney-specific studies, including those related to renal physiology, pathology, and the impact of genetic modifications on kidney function and cellular processes. Overall, the HK-2xZFN-Smc4-mEGFP Clone 82-68 cell line serves as a powerful and versatile model for investigating the fundamental aspects of chromosome biology and kidney cell function, offering insights that are crucial for both basic research and potential clinical applications in nephrology and related fields.
Organism	Human
Tissue	Endocervix
Disease	Adenocarcinoma

Age	30 years
Gender	Female
Ethnicity	African American
Morphology	Epithelial-like cells with mosaic stone shape
Growth properties	Adherent

Citation	HK-2xZFN-Smc4-mEGFP Clone 82-68 (Cytion catalog number 301576)
Biosafety level	1
Depositor	Dr. J. Ellenberg, EMBL Heidelberg

Products	EGFP (Enhanced Green Fluorescent Protein)

Culture Medium	DMEM, w: 4.5 g/L Glucose, w: 4 mM L-Glutamine, w: 1.5 g/L NaHCO3, w: 1.0 mM Sodium pyruvate (Cytion article number 820300a)
Medium supplements	Supplement the medium with 10% FBS
Passaging solution	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.
Split ratio	A ratio of 1:5 in the beginning and up to 1:15 after establishment of the culture is recommended
Fluid renewal	2 to 3 times per week
Freeze medium	CM-1 (Cytion catalog number 800100) or CM-ACF (Cytion catalog number 806100)
Handling of cryopreserved cultures	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. Optionally, skip centrifugation but remove any remaining freezing medium after 24 hours. 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.