HeLa EB3-GFP Kirc 14 09/27/24

HeLa EB3-GFP Kirc 14 is a genetically modified derivative of the human HeLa cervical carcinoma cell line, engineered to stably express a GFP-tagged EB3 (end-binding protein 3) fusion construct, with "Kirc 14" denoting a specific sub-clone (clone 14) selected from the transfected pool. EB3 is a member of the end-binding protein family that tracks growing microtubule plus-ends and serves as a reporter for dynamic microtubule behaviour in living cells. The GFP-EB3 fusion protein produces characteristic comet-like fluorescent signals at polymerizing microtubule tips, enabling live-cell imaging of microtubule dynamics, intracellular transport, and cytoskeletal organisation in real time.

HeLa EB3-GFP Kirc 14 is used in cell biology research for live-cell imaging of microtubule plus-end dynamics, studies of mitotic spindle assembly and kinetochore-microtubule interactions, investigations of intracellular cargo transport, and evaluation of compounds that affect microtubule polymerization dynamics. The stable sub-clonal GFP-EB3 expression provides consistent fluorescence intensity and pattern across passages, making clone 14 suitable for quantitative imaging assays and high-throughput screening of microtubule-targeting agents.

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.