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
Julie DAM-PLAT,Patty CHEN-CHAVAND

Publications:

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).

Reference:

RT00006

Type of research tool: Cell Lines

Therapy area(1): Metabolic Disorders

Therapy area(2): Others

Uses(1): Academic Research

Uses(2): Industry Research (screening, tox.studies, bioreactor, ...)

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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

Tagged leptin receptors hek293t cells

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