生物源
human
重組細胞
expressed in Chem-1 cells
製造商／商標名
ChemiScreen
Chemicon®
技術
radioligand binding assay (RLBA): suitable
NCBI登錄號
UniProt登錄號
運輸包裝
dry ice
一般說明
Full-length human CALCRL and RAMP1 cDNAs encoding CGRP1
The calcitonin family of peptides, which includes calcitonin, two calcitonin-related gene peptides (CGRPs), adrenomedullin, and amylin, share a disulfide-bonded loop, an amphipathic a-helix, and an amidated C-terminus. The peptides influence a variety of physiological effects, including vascular tone, food intake and bone metabolism. The biological effects of the peptides are mediated by two class B GPCRs, CT receptor (calcitonin receptor) and CL receptor (calcitonin receptor-like receptor) (Poyner et al., 2002). The pharmacology of the CT and CL receptors is strongly influenced by their association with a family of three receptor activity modifying proteins (RAMPs), which are 14-17 kD single pass transmembrane proteins. CGRP (calcitonin gene-related peptide) induces vasodilation, and its activity is primarily mediated by CGRP1, composed of CL in complex with RAMP1 (Hasbak et al., 2003). As a consequence of its role as a vasodilator, CGRP acting via CGRP1 appears to play an important role in hypertension (Deng and Li, 2005). Millipore′s CGRP1 membrane preparations are crude membrane preparations made from our proprietary stable recombinant cell lines to ensure high-level of GPCR surface expression; thus, they are ideal HTS tools for screening of antagonists of CGRP1 interactions with CGRP. The membrane preparations exhibit a Kd of 1.1-1.7 nM for [125I]-hCGRP. With 5 µg/well CGRP1 Membrane Prep and 1 nM [125I]-hCGRP, a greater than 15-fold signal-to-background ratio was obtained.
生化／生理作用
GPCR Class: B
Protein Target: CGRP1
Target Sub-Family: Calcitonin
品質
SPECIFICATIONS:
Signal:background and specific binding values obtained in a competition binding assay with varying amounts of CGRP1 membrane prep:
1 unit = 5 µg membrane preparation
Bmax: 10.8 pmol/mg
Kd: 1.4 nM
Signal:background and specific binding values obtained in a competition binding assay with varying amounts of CGRP1 membrane prep:
| 10 µg/well | 5 µg/well | |
|---|---|---|
| Signal:Background | 26.5 | 29.3 |
| Specific Binding (cpm) | 55556 | 47919 |
1 unit = 5 µg membrane preparation
Bmax: 10.8 pmol/mg
Kd: 1.4 nM
規格
Inucbation Conditions
Membranes are mixed with radioactive ligand and unlabeled competitor (see Figures 1 and 2 for concentrations tested) in binding buffer in a nonbinding 96-well plate, and incubated for 1-2 h. Prior to filtration, a GF/C 96-well filter plate is coated with 0.33% polyethyleneimine for 30 min, then washed with 50mM HEPES, pH 7.4, 0.5% BSA. Binding reaction is transferred to the filter plate, and washed 3 times (1 mL per well per wash) with Wash Buffer. The plate is dried and counted.
Binding buffer: 50 mM Hepes, pH 7.4, 5 mM MgCl2, 1 mM CaCl2, 0.2% BSA, filtered and stored at 4°C
Radioligand: [125I] hCGRP (Perkin Elmer # NEX354)
Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl , 0.1% BSA, filtered and stored at 4°C.
Membranes are mixed with radioactive ligand and unlabeled competitor (see Figures 1 and 2 for concentrations tested) in binding buffer in a nonbinding 96-well plate, and incubated for 1-2 h. Prior to filtration, a GF/C 96-well filter plate is coated with 0.33% polyethyleneimine for 30 min, then washed with 50mM HEPES, pH 7.4, 0.5% BSA. Binding reaction is transferred to the filter plate, and washed 3 times (1 mL per well per wash) with Wash Buffer. The plate is dried and counted.
Binding buffer: 50 mM Hepes, pH 7.4, 5 mM MgCl2, 1 mM CaCl2, 0.2% BSA, filtered and stored at 4°C
Radioligand: [125I] hCGRP (Perkin Elmer # NEX354)
Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl , 0.1% BSA, filtered and stored at 4°C.
外觀
One package contains enough membranes for at least 200 assays (units), where an unit is the amount of membrane that will yield greater than 15-fold signal:background with 125I-labeled hCGRP at 1 nM. Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA with no preservatives. Packaging method: Membranes protein were adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80°C.
儲存和穩定性
Store at –70°C. Product is stable for at least 6 months from the date of receipt when stored as directed. Do not freeze and thaw.
法律資訊
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
免責聲明
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.
儲存類別代碼
12 - Non Combustible Liquids
水污染物質分類（WGK）
WGK 2
閃點（°F）
Not applicable
閃點（°C）
Not applicable
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