HeLa EB3 GFP KIRC14 Cells

Name: CLS Cell Lines Service GmbH
Address: Dr.-Eckener-Straße 8, Eppelheim, 69214, DE
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cryovial

Product number: 305718

Safety data sheet

Product sheet

Certificate of Analysis (CoA)

Description	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.
Organism	Human
Tissue	Cervix
Disease	Cervical adenocarcinoma (HeLa background); stably transfected with GFP-EB3 (microtubule plus-end reporter)
Applications	Live-cell microtubule dynamics imaging; mitotic spindle studies; kinetochore-microtubule interaction research; intracellular transport; microtubule-targeting drug screening

Morphology	Epithelial-like
Cell type	Epithelial cells
Growth properties	Adherent

Citation	HeLa EB3-GFP Kirc 14 09/27/24 (Cytion catalog number 305718)
Biosafety level	1
NCBI_TaxID	9606
CellosaurusAccession	Derived from HeLa (CVCL_0030); GFP-EB3 sub-clone not independently assigned
GMO Status	GMO-S1: This HeLa EB3 GFP KIRC14 line contains a GFP-tagged EB3 construct for microtubule plus-end tracking in live-cell imaging. This classification applies only within Germany and may differ elsewhere.

Culture Medium	DMEM, w: 4.5 g/L Glucose, w: 4 mM L-Glutamine, w: 3.7 g/L NaHCO3, w: 1.0 mM Sodium pyruvate (Cytion article number 820300a)
Supplements	Supplement the medium with 10% FBS
Dissociation Reagent	Accutase, 10 min., 37°C
Doubling time	approx. 24 hours
Subculturing	Remove the old medium from the adherent cells and wash with PBS. Cover with Accutase, incubate at room temperature for 8-10 minutes, resuspend with medium, centrifuge at 300×g for 3 minutes, and transfer to fresh flasks with selection medium.
Split ratio	1 to 5
Seeding density	1 to 3 × 10⁴ cells/cm²
Fluid renewal	2 to 3 times per week
Freeze medium	As a cryopreservation medium, we use complete growth medium + 10% DMSO for adequate post-thaw viability.
Thawing and Culturing Cells	Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit. 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. 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. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening. 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. Centrifuge the mixture at 200 x g for 5 minutes, carefully discard the supernatant containing freezing medium. Follow the procedure described under Post-Thaw Recovery
Incubation Atmosphere	37°C, 5% CO₂, humidified atmosphere.
Shipping Conditions	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.
Storage Conditions	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.

Lot Number	Certificate Type	Date	Catalog Number
305718-260526	Certificate of Analysis	13. Jul. 2026	305718

HeLa EB3 GFP KIRC14 Cells

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Certificate of Analysis (CoA)