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HK EGFP-LaminA/H2B-mCherry Cells

HK EGFP-LaminA/H2B-mCherry Cells

Für die Verwendung dieses Produktes fordert der Hersteller das zu Ihrer Institution passende, ausgefüllte und unterschriebene Formular:

Profit Organisation/Nutzung: CLS_CLEAR.pdf
Non-Profit Organisation/Nutzung: Non_Profit_CLS_Supply_Agreement.pdf

Bitte laden Sie dieses entweder beim Kaufabschluss hoch oder senden es alternativ an die purchasing@hoelzel.de

Item no.	CLS-300921
Manufacturer	CLS Cell Lines Service
Amount	1 cryovial
Category	Cell Lines & Cell Culture Cell Lines
Type	Cell line
Certificate	For a certificate of analysis, please send an inquiry to info@hoelzel.de.
Specific against	Human (Homo sapiens)
Dry ice	Yes
ECLASS 10.1	42040401
ECLASS 11.0	42040401
UNSPSC	41106509
Alias	HeLa Kyoto EGFP-LaminA and H2B-mCherry
Available
Manufacturer - Category	Reporter-labeled cells
Description	The HK EGFP-LaminA/H2B-mCherry cell line is a genetically engineered Hela Kyoto derived cell model developed to facilitate advanced studies in nuclear dynamics and chromatin organization within living cells. This cell line expresses two fusion proteins: EGFP (enhanced green fluorescent protein) fused with Lamin A, and mCherry (a red fluorescent protein) fused with Histone H2B. The EGFP-Lamin A fusion highlights the nuclear envelope and allows for the visualization of nuclear architecture changes during cell cycle progression or under various experimental conditions. Meanwhile, the H2B-mCherry fusion protein binds to DNA and provides a vivid red fluorescence that marks chromatin, enabling real-time observation of chromosomal processes during mitosis and interphase. These cells are invaluable for real-time imaging applications, including studies on nuclear integrity, DNA replication, and cellular aging, as well as research into diseases where nuclear architecture is disrupted, such as cancer and laminopathies. The dual-color fluorescence feature of this cell line allows for simultaneous visualization of both the nuclear envelope and chromatin, facilitating a comprehensive understanding of nuclear-cytoplasmic interactions and the spatiotemporal organization of chromatin. Such capabilities make it a critical tool for molecular biology research and cellular biophysics, providing insights into the mechanics of gene expression regulation, nuclear organization, and the cell cycle.
Tissue	Cervix
Growth properties	Monolayer, adherent
Disease	Carcinoma
Age	30 years
Gender	Female
Ethnicity	African American
Morphology	Epithelial-like cells with mosaic stone shape
Biosafety Level	1
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
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.
Fluid Renewal	2 to 3 times per week
Freezing Recovery	After thawing, plate the cells at 5 x 10^4 cells/cm^2 and allow the cells to recover from the freezing process and to adhere for at least 24 hours.
Freeze Medium	CM-1 (Cytion catalog number 800100)
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. 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.
Disclaimer	Our cells are provided for in vitro laboratory research purposes exclusively and are not intended for clinical or diagnostic use, nor are they to be administered to humans or used for veterinary purposes. Users must adhere to all applicable guidelines and regulations for the handling and use of these cells in a research setting.
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.
Split Ratio	A ratio of 1:3 is recommended
Seeding Density	1 x 10^4 cells/cm^2
Products	Histone H2B
Protein Expression	EGFP-LaminA/H2B-mCherry

Note: The presented information and documents (Manual, Product Datasheet, Safety Datasheet and Certificate of Analysis) correspond to our latest update and should serve for orientational purpose only. We do not guarantee the topicality. We would kindly ask you to make a request for specific requirements, if necessary.

All products are intended for research use only (RUO). Not for human, veterinary or therapeutic use.

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CLS-300921-datasheet.pdf

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Amount: 1 cryovial

available

Delivery expected until 10/2/2025

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