CELL LINES
Contributor Information
- Name Alessandra Cambi
- Institute Radboud University Medical Center
Tool Details
- Tool name: CD6-LifeAct-GFP Jurkat cell line cell line
- Alternate names: CD6
- Tool type: Cell Lines
- Tool sub-type: Continuous
- Parental cell line: Jurkat E6.1
- Organism: Human
- Tissue: Blood
- Cancer type: Blood cancer
- Disease: Cancer
- Conditional: No
- Description: Activation of the T cell receptor (TCR) on the T cell through ligation with antigen-MHC complex of an antigen-presenting cell (APC) is an essential process in the activation of T cells and induction of the subsequent adaptive immune response. Upon activation, the TCR, together with its associated co-receptor CD3 complex, assembles in signaling microclusters that are transported to the center of the organizational structure at the T cell-APC interface termed the immunological synapse (IS). During IS formation, local cell surface receptors and associated intracellular molecules are reorganized, ultimately creating the typical bull's eye-shaped pattern of the IS. CD6 is a surface glycoprotein receptor, which has been previously shown to associate with CD3 and co-localize to the center of the IS in static conditions or stable T cell-APC contacts.
- Research area: Cancer; Cell biology; Cell signaling and signal transduction; Immunology
- Production details: Jurkat cell lines stably expressing CD6-LifeAct-YFP were obtained by electroporation using the Neon Transfection System for Electroporation (Invitrogen) according to the manufacturer's guidelines. Shortly, 5*105 Jurkat cells were transfected at 1325 Volt (10 ms, 3 pulses) with 3 ÄÂÄ?g of DNA in 100 ÄÂÄ?l Resuspension buffer. After transfection cells were seeded in 2 ml of 1640 RPMI with 10% FCS and 1% U-glut. Antibiotics were added after 3 h. Stable cell lines were sorted on YFP expression on a FACSAria cell sorter (BD Biosciences), and cells were maintained in complete RPMI medium as described above, additionally supplemented with 500 ng/ml geneticin (G418, Gibco).
- For Research Use Only
Target Details
- Target: Cluster of Differentiation 6 labelled with RFP, CD6-RFP
Application Details
Handling
- Format: Frozen
- Growth medium: 1640 RPMI supplemented with 10% Fetal Calf Serum, 1 mM Ultra-glutamine and antibiotics (100 U/ml penicillin, 100??g/ml streptomycin and 0.25 ?g/ml amphotericin B). 500 ng/ml geneticin.
- Shipping conditions: Dry ice
Documentation
Related Tools
References
- • Meddens et al. 2018. Front Immunol. 9:2333. PMID: 30356797.
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