U2OS-CRISPR-SNAPf-SEH1 Cells

USUSD$800.00*

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).

價格不含稅，另加運費

cryovial

產品編號： 300664

安全資料表

產品說明書

說明	U2OS-CRISPR-SNAPf-SEH1 is a genome-edited human osteosarcoma cell line derived from U2OS cells in which the endogenous SEH1L (SEH1) gene has been modified using CRISPR/Cas9 technology to encode an in-frame SNAPf tag. SEH1 is a component of the Y-complex (also known as the NUP107-160 complex), a core structural module of the nuclear pore complex (NPC) that contributes to pore scaffold assembly and stability. By inserting the SNAPf coding sequence at the endogenous locus, the tagged SEH1 protein is expressed under native regulatory control, preserving physiological expression levels and minimizing perturbations to nuclear pore composition. The SNAPf tag is an engineered, fast-reacting variant of the SNAP-tag that covalently binds benzylguanine-conjugated substrates, allowing selective and stable fluorescent labeling in live or fixed cells. In U2OS-CRISPR-SNAPf-SEH1 cells, the fusion protein localizes to the nuclear envelope in a punctate pattern characteristic of NPC distribution. Because labeling occurs at endogenous protein levels, this system is well suited for quantitative fluorescence microscopy, super-resolution imaging, and single-particle tracking analyses aimed at dissecting NPC organization and stoichiometry. The flat morphology and large nuclei of U2OS cells further facilitate high-resolution visualization of nuclear envelope structures. SEH1 participates in NPC biogenesis and has also been implicated in kinetochore-associated processes during mitosis. Accordingly, this cell line provides a robust platform for investigating cell cycle-dependent NPC assembly and disassembly, spatial organization of the Y-complex within the pore scaffold, and potential dual roles of SEH1 at the nuclear envelope and mitotic kinetochores. U2OS-CRISPR-SNAPf-SEH1 enables mechanistic studies of nuclear pore architecture and dynamics under physiologically relevant expression conditions.
生物體	Human
組織	Bone
疾病	Osteosarcoma

年齡	15 years
性別	Female
族裔	Caucasian
形態學	Epithelial-like
生長特性	Adherent

引用	U2OS-CRISPR-SNAPf-SEH1 (Cytion catalog number 300664)
生物安全等級	1
NCBI_TaxID	9606
存款人	The Ellenberg Lab (EMBL)
轉基因狀態	GMO-S1: This human osteosarcoma cell line (U2OS-CRISPR-SNAPf-SEH1) contains a CRISPR-mediated SNAPf-SEH1 fusion that allows selective labeling of the SEH1 nucleoporin. The modification is stably present. This classification applies only within Germany and may differ elsewhere.

蛋白質表達	SEH1, SNAPf-tag

培養基	McCoys 5a, w: 3.0 g/L Glucose, w: stable Glutamine, w: 2.0 mM Sodium pyruvate, w: 2.2 g/L NaHCO3 (Cytion article number 820200a)
營養補充品	Supplement the medium with 10% FBS, 3.0 g/L Glucose, stable Glutamine, 2.0 mM Sodium pyruvate, 2.2 g/L NaHCO3, 1% NEAA
解離試劑	Accutase
傳代培養	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.
冷凍培養基	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.
細胞解凍與培養	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.
培養環境	37°C, 5% CO₂, humidified atmosphere.
運送條款	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.
儲存條件	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.

不孕症	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.

U2OS-CRISPR-SNAPf-SEH1 Cells

一般資訊

特徵

監管數據

生物分子資料

處理方式

品質控制與分子分析