NRK-EGFP-H2B

The NRK-EGFP-H2B cell line is a genetically modified variant of normal rat kidney (NRK) cells that stably express the enhanced green fluorescent protein (EGFP) fused to histone H2B. This modification allows for real-time visualization of chromatin and nuclear dynamics, making this cell line an invaluable tool for studying cell cycle progression, mitosis, and chromatin organization. The stable expression of EGFP-H2B provides a bright and consistent fluorescent signal, facilitating high-resolution live-cell imaging and enabling researchers to monitor nuclear events with great precision.

NRK cells, originating from the kidney tissue of an adult rat, are widely used in cellular biology due to their robust growth characteristics and well-documented physiological behaviors. The introduction of the EGFP-H2B fusion protein into these cells does not significantly alter their growth or morphology, allowing for reliable and reproducible experimental conditions. This cell line is particularly useful in studies of kidney cell biology, cellular responses to stress, and mechanisms of carcinogenesis, given the kidney's role in filtering blood and excreting waste. Additionally, the fluorescence capabilities of NRK-EGFP-H2B cells can be harnessed in drug screening applications to observe drug effects on cell proliferation and nuclear morphology in real-time.

37°C, 5% CO₂, humidified atmosphere.

1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

2. 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.

3. 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.

4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

5. 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.

6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

7. 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.

8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

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.

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.

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.

At Cytion, we stand behind the quality and viability of our products. We guarantee that our cells will be viable at the start of culture for 60 days after shipment from our facilities. We assure that all materials conform to the specifications described in the Certificate of Analysis and/or Product Sheet.

For full details, please refer to our Warranty Statement.