Stable HEK293 Cells-Rpn11-HTBH

Stable HEK293 Cells-Rpn11-HTBH is a stably transfected derivative of the HEK293 (Human Embryonic Kidney 293) cell line, engineered to express a tagged version of Rpn11 (also known as PSMD14 or POH1), the deubiquitinase subunit of the 26S proteasome lid complex. Rpn11 is a Zn²⁺-dependent JAMM-domain deubiquitinase that removes ubiquitin chains from proteasome-bound substrates during proteasomal degradation. The HTBH tag (hexahistidine-TEV-biotin acceptor peptide-hexahistidine) enables affinity purification of Rpn11-containing complexes under native conditions, making this line particularly suited for proteasome complex purification and interactome studies.

This cell line is applicable in studies of 26S proteasome biology, ubiquitin-proteasome pathway (UPS) regulation, Rpn11/PSMD14 function in protein quality control, proteasome assembly and dynamics, and proteasome inhibitor mechanism of action. It is also used for affinity purification of native proteasome complexes and as a model for studying deubiquitinase biology in the context of the proteasome. The HTBH tagging system enables highly stringent purification of biotinylated complexes using streptavidin-based pulldowns.

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 200 x g for 5 minutes, carefully discard the supernatant containing freezing medium.

7. Follow the procedure described under Post-Thaw Recovery

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.