ChemiScreen™ Human Recombinant BB3 Bombesin Receptor Calcium-Optimized Stable Cell Line

ChemiScreen™ Human Recombinant BB3 Bombesin Receptor Calcium-Optimized Stable Cell Line

• ChemiScreen
• Chemicon (Millipore)

2 vials

Full-length human BRS3 cDNA encoding BB3.

Bombesin, a bioactive peptide first identified in amphibian skin, is related to two mammalian peptides, gastrin-releasing peptide (GRP) and neuromedin B. A family of 3 GPCRs, including GRP-R (BB1), NMB-R (BB2) and BRS-3 (BB3), mediate the biological effects of the peptides (Ohki-Hamazaki et al., 2005). BB3 differs from the others by its low affinity for bombesin. Although an endogenous ligand for BB3 has yet to be identified, a synthetic nonselective bombesin-like peptide [D-Phe⁶, β-Ala¹¹, Phe¹³, Nle¹⁴]-bombesin-(6-14)-amide activates BB3 with high potency. BB3-null mice develop mild obesity that can be alleviated by treatment with an antiobesity compound, the serotonin and noradrenaline reuptake inhibitor Sibutramine (Matsumoto and Iijima, 2003). Chemicon's cloned human BB3-expressing cell line is made in the Chem-1 host, which supports high levels of recombinant BB3 expression on the cell surface and contains high levels of the promiscuous G protein Gα15 to enhance coupling of the receptor to the calcium signaling pathway. Thus, the cell line is an ideal tool for screening for agonists and antagonists at BB3.

Calcium flux assay, ligand binding assays

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Mammalian bombesin-like peptides (see MIM 137260) are widely distributed in the central nervous system as well as in the gastrointestinal tract, where they modulate smooth-muscle contraction, exocrine and endocrine processes, metabolism, and behavior. They bind to G protein-coupled receptors on the cell surface to elicit their effects. Bombesin-like peptide receptors include gastrin-releasing peptide receptor (MIM 305670), neuromedin B receptor (MIM 162341), and bombesin-like receptor-3 (BRS3) (Ohki-Hamazaki et al., 1997).[supplied by OMIM]

FUNCTION: SwissProt: P32247 # Role in sperm cell division, maturation, or function. This receptor mediates its action by association with G proteins that activate a phosphatidylinositol-calcium second messenger system.
SIZE: 399 amino acids; 44411 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: In germ cells in testis. Lung carcinoma cells.
SIMILARITY: SwissProt: P32247 ## Belongs to the G-protein coupled receptor 1 family.

Human

SPECIFICATIONS: EC50 for calcium mobilization by [D-Phe⁶, β-Ala⁶, Phe¹³, Nle¹⁴]-Bombesin-(6-14)-amide: ~ 1.35 nM

EC50 for calcium mobilization by Bombesin: >100 nM

Chem-1

GPCR Cell Lines

Chemicon

Chem-1

Cells are frozen at 2 x 10⁶ cells/mL in Chemicon Cell Culture Freezing Media with DMSO. Cell line tests negative for mycoplasma.

BB3

P32247

peptide

Place cells in liquid nitrogen immediately upon receipt. Maintain frozen in liquid nitrogen for up to 5 years. Thaw cells rapidly by removing from liquid nitrogen and immediately immersing in a 37ºC water bath. Immediately after ice has thawed, sterilize the exterior of the vial with 70% ethanol. Transfer contents of the vial to a T75 flask containing 20 mL growth media, and place in a humidified 37°C incubator with 5% CO2. After 8-24 h, cells will adhere to the plate, at which time the media should be replaced to remove residual DMSO. Cells are passaged by washing with Ca⁺⁺ and Mg⁺⁺-free HBSS (10 mL/T75), incubating with 0.05% trypsin/0.2 g/L EDTA (1 mL/T75) for 5-10 minutes at 37°C, and rapping the side of the flask to dislodge the cells. Neutralize the trypsin by addition of 4 volumes growth media. Cells are typically passaged 1:10 with every 3-4 days, and should be passaged at least once after thawing prior to use in calcium flux assays.

Bombesin

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A

2 x 10⁶ cells/vial

• BRS3
• BRS-3

GPCRs

ChemiScreen™ Human Recombinant BB3 Bombesin Receptor Calcium-Optimized Stable Cell Line

NM_001727.1

DMEM containing 4.5 g/L glucose/10% heat inactivated fetal bovine serum/1x nonessential amino acids/10 mM HEPES/0.25 mg/ml Geneticin (G418)/100 U/ml each penicillin and streptomycin.