Tagged leptin receptors hek293t cells

Hek293T cells were transfected with short form human leptin receptors (OBR) tagged individually with either Rluc (pIRESpuro3, Clontech) or YFP (pIEShyg3, Clontech). After antibiotic selection (puromycin and hygromycin), single cell clones were isolated and cultured, then analyzed by microscopy, luminescence measures, western blot and BRET.

Interest / Relevance: The cell line may be used to screen for OBR ligands (agonists, antagonists, competitive or allosteric. These ligands may be usefull for the treatment of OBR-associated diseases such as obesity, anorexia, auto-immune diseases etc.rnCells expressing only one type of tagged OBR (either Rluc or YFP) can be used to quantify OBR expression either by fluorescence or luminescence.
Scientist's name: Ralf JOCKERS
Couturier C. and Jockers R.: Activation of the leptin receptor by a ligand-induced conformational change of constitutive receptor dimers. J. Biol. Chem. 278: 26604-26611 (2003).



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Organism: Human, Homo sapiens
Tissue: Kidney
Morphology: Epithelial
Passage Number: 16
Subculture Routine: Protocol for 90 mm dish: 1) Remove and discard culture medium. 2) Briefly rinse cell layer with PBS to remove serum traces. 3) Add 2ml Trypsin-EDTA and observe under microscope until cell layer is dispersed, usually within 5-10 minutes. 4) Add 6ml complet
Culture Medium: Delbeccos Modified Eagles Medium (Gibco, ) with 10% heat inactivated Fetal Calf Serum.
Growth Properties: Cells adhere to substrate and grow as a monolayer when incubated at 37°C. Cells detach and remain in suspension if left at room temperature for a considerable amount of time. The cells will re-attach to the substrate over a period of several hours under c
Karyotype: null
Rare disease: No
Last update: 12/06/2024

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