NRK-Pom121-EGFP3

The NRK-Pom121-EGFP3 cell line is derived from normal rat kidney (NRK) cells and is genetically engineered to express the Pom121-EGFP3 fusion protein. Pom121 is a transmembrane nucleoporin that is an integral component of the nuclear pore complex (NPC), playing a crucial role in nuclear envelope assembly and NPC function. The inclusion of the enhanced green fluorescent protein (EGFP3) tag facilitates the visualization and study of Pom121 dynamics, localization, and interactions within live cells through fluorescence microscopy. This makes the NRK-Pom121-EGFP3 cell line a valuable tool for investigating nuclear transport mechanisms and NPC architecture.

NRK cells, the parental line of NRK-Pom121-EGFP3, are commonly used in various research applications due to their stable growth characteristics and epithelial morphology. The modification to express Pom121-EGFP3 provides researchers with a robust model to examine the molecular mechanisms underlying nucleocytoplasmic transport, the structural organization of the NPC, and its regulation during cell division and differentiation. Additionally, this cell line can be used to study the effects of various genetic and pharmacological perturbations on NPC function, offering insights into diseases associated with nuclear transport defects, such as cancer and neurodegenerative disorders.

Overall, the NRK-Pom121-EGFP3 cell line represents a sophisticated tool in cell biology and molecular research, providing high-resolution insights into the dynamic processes governing nucleocytoplasmic interactions. Its ability to allow real-time observation of NPC components in a live cellular context makes it invaluable for advancing our understanding of cellular transport mechanisms and their implications in health and disease.

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.