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CD6-RFP Jurkat cell line

Invented by Alessandra Cambi
Invented at Radboud UMC

Info

Catalogue Number 160713
Antigen/Gene or Protein Targets Cluster of Differentiation 6 labelled with RFP, CD6-RFP
Parental Line Jurkat E6.1
Synonyms CD6
Host Human
Tissue Blood
Disease Keywords Acute T cell Leukemia
Relevance 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.
Production Details Jurkat cell lines stably expressing CD6-RFP 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 μg of DNA in 100 μl 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 RFP 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).
Conditional No
Research Area Adhesion, Cancer, Cell Signaling & Signal Transduction, Immunology
Recommended Growing Conditions 1640 RPMI (PAA) supplemented with 10% Fetal Calf Serum, 1 mM Ultra-glutamine and antibiotics (100 U/ml penicillin, 100 μg/ml streptomycin and 0.25 μg/ml amphotericin B). 500 ng/ml geneticin.

References

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References: 1 entry

Meddens et al. 2018. Front Immunol. 9:2333. PMID: 30356797.


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References: 1 entry

Meddens et al. 2018. Front Immunol. 9:2333. PMID: 30356797.


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Inventor Information

Inventors

Alessandra  Cambi

Alessandra Cambi

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