HTS040M
ChemiSCREEN Membrane Preparation Recombinant Human NOP Opioid Receptor
Human NOP / ORL1 / OP4 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
About This Item
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General description
Application
Biochem/physiol Actions
Quality
5 µg/well | |
---|---|
Signal:Background | 23.8 |
Specific Binding (cpm) | 6656 |
SPECIFICATIONS: 1 unit = 5 µg
Bmax for [125I]-Nociceptin binding: 1.1 pmol/mg protein
Kd for [125I]-Nociceptin binding: ~0.06 nM
Specifications
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 50 mM Tris-HCl, pH 7.4. 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 Tris-HCl, pH 7.4 filtered and stored at 4°C
Radioligand: [125I]-Nociceptin. (Perkin Elmer#:NEX-324 )
Wash Buffer: 50 mM Hepes, pH 7.4, 500 mM NaCl , 0.1% BSA, filtered and stored at 4°C.
One package contains enough membranes for at least 200 assays (units), where a unit is the amount of membrane that will yield greater than 10-fold signal:background with 125I labeled Nociceptin
Physical form
Packaging method: Membranes protein were adjusted to the indicated concentration in 1 ml 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
flash_point_f
Not applicable
flash_point_c
Not applicable
Certificates of Analysis (COA)
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