HeLa EB3 GFP KIRC14 Cells
General information
| 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 |
Characteristics
| Morphology | Epithelial-like |
|---|---|
| Cell type | Epithelial cells |
| Growth properties | Adherent |
Regulatory Data
| 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. |
Biomolecular Data
Handling
| 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 |
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| 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. |
Quality Control & Molecular Analysis
Certificate of Analysis (CoA)
| Lot Number | Certificate Type | Date | Catalog Number |
|---|---|---|---|
| 305718-260526 | Certificate of Analysis | 13. Jul. 2026 | 305718 |
Material Transfer Agreement
If you intend to use Cytion cell lines solely for internal research at a single research site, please complete and sign our Material Transfer Agreement (MTA) and submit it along with your order.
For any commercial applications - including but not limited to fee-for-service work, quality control testing, product release, diagnostic use, or regulatory studies - please complete the Intended Use Form so we can prepare a suitable agreement tailored to your project.
Please note: The MTA applies only to certain cell lines. If this notice and the MTA document appear on a product page, the agreement is applicable. For cell lines not covered by the MTA, no reference to the agreement will be shown. The MTA is not valid for customers in the Americas, China, or Taiwan. Please contact our U.S. entity to receive the appropriate agreement.
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Required products
Required products
Freeze Medium CM-1 - 50 mlCryopreservation Media Variants: 50 mlCytion’s Freeze Medium CM-1 is a state-of-the-art cryopreservation medium designed to ensure the highest level of cell viability and functionality post-thaw. This versatile medium is suitable for a broad spectrum of cell types, including both human and animal cells, making it an essential tool for diverse research applications. Formulated with a meticulously balanced combination of cryoprotectants and essential nutrients, Freeze Medium CM-1 minimizes ice crystal formation and cellular stress during the freezing process, thus preserving cellular integrity.
Key features of Freeze Medium CM-1 include:
Broad Compatibility: Effective for a wide range of cell types, including primary cells, stem cells, and established cell lines.
High Viability: Optimized to maximize post-thaw cell recovery and viability, ensuring reliable experimental outcomes.
Ready-to-Use: Conveniently prepared and sterilized for immediate application, reducing preparation time and risk of contamination.
Enhanced Stability: Maintains consistent performance under standard cryopreservation conditions, ensuring reproducible results.
Long Shelf Life: CM-1 is a serum-containing, ready-to-use cryopreservation medium that can be stored in the refrigerator for up to one year.
Using CM-1 for Freezing Cells
To use CM-1 for freezing both adherent and suspension cells, follow these steps:
For adherent cells, wash and dissociate them from the culture substrate. For suspension cells, proceed directly to the next step.
Count the cells to ensure they are at the proper concentration.
Centrifuge the cells to pellet them, then resuspend in CM-1 freeze medium.
Transfer the resuspended cells into cryovials.
Use a slow-freezing method before transferring the cells to long-term storage.
Method
Description
Steps
❄️
Manual Freezing
A step-by-step method involving gradual temperature reduction to ensure cell viability.
1️⃣ Place cells in freeze medium in a 4°C freezer for 40 minutes.
2️⃣ Transfer to a -80°C freezer for 24 hours.
3️⃣ Store cells in liquid nitrogen for long-term preservation.
❄️
Using Mr. Frosty
A convenient device that allows for controlled freezing rates without electrical power.
1️⃣ Prepare cells in cryovials with freeze medium.
2️⃣ Place cryovials in Mr. Frosty container.
3️⃣ Store at -80°C for 24 hours before transferring to liquid nitrogen.
❄️
Controlled-Rate Freezer
A high-precision freezer by Thermo Fisher or other manufacturers designed for controlled temperature reduction.
1️⃣ Program the device to gradually decrease the temperature.
2️⃣ Place prepared cells in the freezer.
3️⃣ After the freezing cycle, transfer cells to liquid nitrogen.
Store the cryovials at temperatures below -130°C or in liquid nitrogen for long-term preservation.
Ingredients
Contains FBS, DMSO, Glucose, Salts
Buffering capacity: pH = 7.2 to 7.6
Cytion’s Freeze Medium CM-1 offers a reliable solution for cryopreservation, ensuring high cell viability and functionality post-thaw for a wide range of research applications.€59.00*Accutase Cell Detachment Solution - 100 mlVariants: 100 mlAccutase Cell Detachment Solution with EDTA and Phenol Red – 100 ml
Accutase is a ready-to-use, sterile-filtered cell detachment solution designed as a gentle alternative to trypsin/EDTA for dissociating adherent mammalian cells from standard tissue culture plasticware and adhesion-coated surfaces. It combines proteolytic and collagenolytic enzyme activity in a balanced salt solution to deliver effective yet controlled dissociation, preserving cell-surface proteins and supporting high post-passage viability and rapid reattachment.
The Accutase formulation is based on Dulbecco’s phosphate-buffered saline (DPBS) with EDTA and phenol red as a visual pH indicator. The enzymes are of non-mammalian and non-bacterial origin, making Accutase particularly well suited to stem cell research, vaccine workflows, and any application where animal
- or microbially-derived contaminants must be minimised. The solution auto-inhibits at 37 °C, so no neutralising reagent or serum-containing medium is required after detachment – cells can be transferred directly into fresh medium.
Key Features
Ready-to-use 1x sterile-filtered liquid – no dilution or reconstitution required
Combined proteolytic and collagenolytic enzyme activity for gentle dissociation
Each batch standardised to a defined dissociation activity for lot-to-lot consistency
Non-mammalian and non-bacterial enzyme origin
Auto-inhibits at 37 °C – no neutralising solution needed
Formulated in Dulbecco’s PBS with EDTA
Phenol red included as visual pH indicator
pH 6.8 – 7.8
Typical Applications
Accutase gently dissociates a wide variety of adherent and sensitive cell types, including human embryonic stem cells (hESCs), human induced pluripotent stem cells (iPSCs), neural stem cells, primary neurons, and routinely cultured adherent lines such as HeLa, HEK 293, CHO, MDCK, Vero, NIH/3T3, BHK-21 and A549. Typical use cases include:
Routine subculture and passaging of adherent mammalian cells
Gentle single-cell dissociation of hESCs, iPSCs and other sensitive lines
Sample preparation for flow cytometry and FACS analysis
Analysis of cell-surface markers where epitope integrity matters
Cell migration, proliferation and apoptosis assays
Quiescence assays by serum starvation and oncogene transfection studies
Tumor cell and neural crest cell migration assays
Production scale-up in bioreactor workflows
For routine work, apply approximately 10 ml of Accutase per 75 cm2 of culture surface and incubate for 5–10 minutes at room temperature. The optimal incubation time should be determined for each cell line and should not exceed one hour. Prior to addition, rinse the cell layer with a Ca2+/Mg2+-free salt solution such as DPBS without calcium and magnesium to remove residual serum and divalent cations.
Handling & Storage
Store the unopened bottle frozen at -15 °C or below. Thaw either at room temperature or overnight at +2 °C to +8 °C. Do not thaw Accutase in a 37 °C water bath, as elevated temperatures reduce enzyme activity. After thawing, the solution can be stored for up to 2 months at +2 °C to +8 °C; do not store at room temperature. Do not pre-warm the reagent to 37 °C before application – add it directly to washed cells at room temperature. For long-term shelf life, single-use aliquoting is recommended to avoid repeated thaw cycles. Always work under aseptic conditions.
Quality
Manufactured under strict quality standards. Each batch of Accutase is sterile-filtered and tested for sterility, pH, appearance and dissociation activity to ensure consistent, reproducible performance from lot to lot.
Product Specifications
Specification
Detail
Product typeCell detachment / dissociation reagent
FormatSterile-filtered liquid, ready-to-use
Volume100 ml
Working concentration1x (ready-to-use)
Enzyme activityCombined proteolytic and collagenolytic
Enzyme originNon-mammalian and non-bacterial
Buffer systemDulbecco’s PBS with EDTA
pH indicatorPhenol red
pH range6.8 – 7.8
AppearanceClear, pale red to orange solution
Storage temperature-15 °C or below
Stability after thawingUp to 2 months at +2 °C to +8 °C
Recommended use volume~10 ml per 75 cm2 culture surface
Typical incubation time5 – 10 minutes at room temperature
Shipping conditionsFrozen on dry ice
Intended useFor research use and further manufacturing only
Formulation (Composition per Liter)
Component
Concentration (mg/L)
Inorganic Salts
Sodium chloride (NaCl)8000.00
Disodium hydrogen phosphate (Na2HPO4)1150.00
Potassium chloride (KCl)200.00
Potassium dihydrogen phosphate (KH2PO4)200.00
Other Components
EDTA · 4Na (tetrasodium EDTA)220.00
Phenol red3.00
Proprietary enzyme blend (proteolytic and collagenolytic activity)1x
Accutase is a registered trademark of Innovative Cell Technologies, Inc.€75.00*Antibiotic/Antimycotic Solution (100x)Product Overview
Volume: 100 ml
Storage: ≤-15°C
Sterility: Sterile-filtered
Antibiotic/Antimycotic Solution (100x) is a sterile, ready-to-use concentrate designed to reduce microbial contamination risks in cell culture and related laboratory applications. This 100x solution contains a well-established combination of penicillin, streptomycin, and amphotericin B—providing broad-spectrum antimicrobial activity against Gram-positive and Gram-negative bacteria, yeasts, and filamentous fungi. The formulation is suitable for use in eukaryotic cell cultures, bacterial media, and other contamination-sensitive systems, supporting clean and consistent lab operations.
Application and Benefits Optimized for routine research protocols, this solution is widely used to maintain aseptic conditions in cell culture workflows. It offers reliable performance in contamination-sensitive environments, helping researchers reduce the risk of microbial overgrowth without compromising cell health or experimental reproducibility. The sterile-filtered formulation eliminates the need for additional solubilization steps, supporting streamlined media preparation and reducing variability in daily lab procedures.
Usage and Compatibility To achieve standard working concentrations, dilute the solution 1:100 into your complete culture medium. The product is compatible with a broad range of mammalian cell lines and basal media. With consistent stock availability, researchers benefit from dependable supply continuity and simplified logistics planning. The solution should be stored at ≤ –15 °C and protected from repeated freeze-thaw cycles to maintain stability. For research use only. Not for use in diagnostic or therapeutic procedures. Not for use in humans or animals.€45.00*