NRK-EGFP3-Seh1

The NRK-EGFP3-Seh1 cell line is a clonal stable line derived from normal rat kidney (NRK) cells. This cell line was generated through the transfection of a circular plasmid encoding the EGFP3-Seh1 fusion protein. Following transfection, cells were selected for drug resistance, ensuring the establishment of a stable population expressing the desired construct.

Approximately 50% of the cells in this population express EGFP3-Seh1, a fusion protein combining enhanced green fluorescent protein (EGFP) with Seh1, a protein component of the nuclear pore complex. The presence of EGFP facilitates visualization and tracking of the fusion protein within the cells, enabling researchers to study the dynamics and function of Seh1 in various cellular processes. However, the expression of EGFP3-Seh1 in this cell line exhibits some variegation, indicating variability in expression levels among individual cells within the population.

This cell line is particularly useful for studies involving nuclear pore complex assembly, nucleocytoplasmic transport, and the role of Seh1 in these processes. The fluorescence provided by EGFP allows for live-cell imaging and real-time analysis of protein localization and interactions, making NRK-EGFP3-Seh1 a valuable tool for cell biology and molecular research.

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.