Panoply™ Human GCGR Over-expressing Stable Cell Line

Name: Panoply™ Human GCGR Over-expressing Stable Cell Line
SKU: CSC-SC006160
Availability: InStock

For research use only. Not intended for any clinical use.

Cat. No. : CSC-SC006160

Host Cell : HEK293 (CHO and other cell types are also available) Size : >1x10⁶ frozen cells/vial

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Cell Line Information

Cell Culture Information

Safety and Packaging

Gene Information

Cat. No.	CSC-SC006160
Description	Using Creative Biogene's proprietary lentiviral vectors, we subclone the target gene into lentivector, generate the lentivirus particles, sequentially infect the cell line HEK293 (other cell types are also available according to your requirements), and select the clones constantly expressing target gene at high level.
Target Gene	GCGR
Gene Species	Homo sapiens (Human)
Host Cell	HEK293 (CHO and other cell types are also available)
Host Cell Species	Species varies
Applications	1. Gene expression studies 2. Signaling pathway research 3. Drug screening and toxicology 4. Disease research
Size	2 × 10^6 cells / vial
Stability	Validated for at least 10 passages
Quality Control	Negative for bacteria, yeast, fungi and mycoplasma.
Storage	Liquid nitrogen
Shipping	Dry Ice

Revival

Rapidly thaw cells in a 37°C water bath. Transfer contents into a tube containing pre-warmed media. Centrifuge cells and seed into a 25 cm2 flask containing pre-warmed media.

Mycoplasma	Negative
Format	One frozen vial containing millions of cells
Storage	Liquid nitrogen
Safety Considerations	The following safety precautions should be observed. 1. Use pipette aids to prevent ingestion and keep aerosols down to a minimum. 2. No eating, drinking or smoking while handling the stable line. 3. Wash hands after handling the stable line and before leaving the lab. 4. Decontaminate work surface with disinfectant or 70% ethanol before and after working with stable cells. 5. All waste should be considered hazardous. 6. Dispose of all liquid waste after each experiment and treat with bleach.
Ship	Dry ice

Gene Name	GCGR glucagon receptor [ Homo sapiens ]
Gene Symbol	GCGR
Synonyms	GCGR; glucagon receptor; GGR; GL-R; FLJ97182; MGC138246;
Gene ID	2642
Uni Prot ID	P47871
Chromosome Location	17q25
Function	glucagon receptor activity; guanyl-nucleotide exchange factor activity; peptide hormone binding; receptor activity; signal transducer activity;
Pathway	Class B/2 (Secretin family receptors), organism-specific biosystem; G alpha (q) signalling events, organism-specific biosystem; G alpha (s) signalling events, organism-specific biosystem; GPCR downstream signaling, organism-specific biosystem; GPCR ligand binding, organism-specific biosystem; GPCRs, Class B Secretin-like, organism-specific biosystem; Glucagon signaling in metabolic regulation, organism-specific biosystem;
MIM	138033

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Endogenous satiety hormones offer an attractive target for anti-obesity drugs. Glucagon promotes weight loss by reducing food intake and increasing energy expenditure. To further understand the cellular mechanisms by which glucagon and its related ligands activate the glucagon receptor (GCGR), researchers investigated the interaction between GCGR and receptor activity-modifying protein (RAMP) 2 (a member of the receptor activity-modifying protein family). They investigated the effects of RAMP2 on GCGR using a combination of competition binding experiments, cell-surface enzyme-linked immunosorbent assays, functional assays assessing Gαs and Gαq pathways and β-arrestin recruitment, and small interfering RNA knockdown. This study demonstrated that co-expression of RAMP2 with GCGR reduced GCGR cell surface expression. Confocal microscopy confirmed this, demonstrating that RAMP2 colocalized with GCGR and led to a significant intracellular redistribution of GCGR. Furthermore, the presence of RAMP2 impaired signaling in the Gαs and Gαq pathways, as well as β-arrestin recruitment. This study suggests that RAMP2 may alter the agonist activity and trafficking of GCGR, which may be relevant for the generation of novel peptide analogs with selective agonist activity.

To investigate whether the RAMP2-induced changes in GCGR cAMP accumulation apply to other cell types, the researchers measured cAMP accumulation after glucagon stimulation in human hepatoma 7 cells overexpressing the human GCGR (Huh7-GCGR) with or without RAMP2 knockdown. Huh7-GCGR cells have low levels of endogenous RAMP2 expression, and RAMP2 silencing resulted in approximately 70% RAMP2 knockdown. In Huh7-GCGR cells with RAMP2 knockdown, there was no statistically significant change in glucagon titer, and a trend toward decreased Emax was observed, but this was not statistically significant (Figure 1).

Figure 1. In Huh7-GCGR cells, the knockdown of RAMP2 resulted in no change in potency and a trend toward decreased efficacy. (Cegla J, et al., 2017)

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