HK-CRISPR-Nup188-mEGFP Cells

Name: CLS Cell Lines Service GmbH
Address: Dr.-Eckener-Straße 8, Eppelheim, 69214, DE
Telephone: +496221405780
Price range: €€€

1 104,00 CAD$*

Products are shipped frozen on dry ice in cryotubes. Each cryotube typically contains 3 × 10⁶ cells for adherent lines or 5 × 10⁶ cells for suspension lines (refer to the batch CoA for details).

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cryovial

Numéro de produit : 300657

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Certificat d'analyse (CoA)

Description	HK-CRISPR-Nup188-mEGFP is a genetically modified Hela Kyoto cell line. It uses CRISPR/Cas9 technology to integrate mEGFP (monomeric Enhanced Green Fluorescent Protein) at the Nup188 locus. Nup188 is a key nucleoporin protein involved in the nuclear pore complex, essential for molecule transport between the nucleus and cytoplasm. The mEGFP tagging allows real-time visualization of Nup188, aiding in the study of nuclear pore complex dynamics and function. This cell line is particularly useful for researchers focusing on nuclear transport and nuclear pore complex biology. Its fluorescent tagging enables observation of Nup188 under various conditions, including drug treatments and genetic modifications. Applications include high-content screening, live-cell imaging, and other fluorescence-based techniques, providing valuable insights into nucleocytoplasmic transport and nuclear envelope integrity.
Organisme	Human
Tissu	Endocervix
Maladie	Adenocarcinoma
Site métastatique	Not applicable (HeLa Kyoto derivative; primary tumor site is endocervix)
Applications	Nuclear pore complex (NPC) biology; nucleocytoplasmic transport; Nup188 dynamics and function; live-cell fluorescence imaging of nuclear envelope; high-content screening; CRISPR knock-in validation studies; nuclear transport inhibitor evaluation

Âge	30 years
Genre	Female
Origine ethnique	African American
Morphologie	Epithelial-like cells with mosaic stone shape
Type de cellule	Epithelial cells
Propriétés de croissance	Adherent

Référence	HK-CRISPR-Nup188-mEGFP (Cytion catalog number 300657)
Niveau de biosécurité	1
NCBI_Numéro d'identification fiscale	9606
Cellosaurus - Numéro d'enregistrement	Not assigned (HK-CRISPR-Nup188-mEGFP is a CRISPR-modified HeLa Kyoto derivative; parental HeLa Kyoto CVCL_1922)
Déposant	The Ellenberg Lab (EMBL)
Statut OGM	GMO-S1: This HeLa Kyoto line contains a CRISPR knock-in of mEGFP at the Nup188 locus, enabling visualization of nuclear pore scaffold dynamics. This classification applies only within Germany and may differ elsewhere.

Expression des protéines	Nup188, mEGFP-tag

Milieu de culture	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)
Suppléments	Supplement the medium with 10% FBS
Réactif de dissociation	Accutase
Repiquage	Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium.
Milieu de congélation	As a cryopreservation medium, we use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress.
Décongélation et culture des cellules	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 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium. 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. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.
Atmosphère d'incubation	37°C, 5% CO₂, humidified atmosphere.
Conditions d'expédition	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.
Conditions d'entreposage	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.

Stérilité	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.

Numéro de lot	Type de certificat	Date	Numéro de catalogue
300657-171123	Certificat d'analyse	23. May. 2025	300657

HK-CRISPR-Nup188-mEGFP Cells

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Certificat d'analyse (CoA)