HTS046M
ChemiSCREEN Membrane Preparation Recombinant Human ETB Endothelin Receptor
Human ETB GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
About This Item
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General description
Application
Biochem/physiol Actions
Quality
10 µg/well | |
---|---|
Signal:Background | 7.4 |
Specific Binding (cpm) | 5704 |
SPECIFICATIONS: 1 unit = 10 μg membrane preparation
Bmax: 0.49 pmol/mg
Kd: 0.03-0.06 nM
Specifications
RECOMMENDED ASSAY 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, an FC 96-well harvest plate (Millipore cat. # MAHF C1H) 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]Endothelin-1 (Perkin Elmer # NEX259)
Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl , 0.1% BSA, filtered and stored at 4°C.
One package contains enough membranes for at least 200 assays (units), where an unit is the amount of membrane that will yield greater than 5-fold signal:background with 125I-labeled Endothelin-1 at 0.25 nM.
Physical form
Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA with no preservatives.
Packaging method: Membrane protein was adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80°C.
Storage and Stability
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Storage Class
12 - Non Combustible Liquids
wgk_germany
WGK 2
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