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Key Documents

HTS056M

Sigma-Aldrich

ChemiSCREEN Membrane Preparation Recombinant Human FPRL1 (Formyl Peptide Receptor-Like 1)

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About This Item

UNSPSC Code:
41123201
eCl@ss:
32161000
NACRES:
NA.45

description

GPCR Class A
protein target FPRL1 / ALX / HM63

Quality Level

concentration

0.5 mg/mL

technique(s)

radioligand binding assay (RLBA): suitable

UniProt accession no.

Gene Information

human ... FPR2(2358)

General description

FUNCTION: SwissProt: P25090 # Low affinity receptor to N-formyl-methionyl peptides, which are powerful neutrophils chemotactic factors. Binding of FMLP to the receptor causes activation of neutrophils. This response is mediated via a G-protein that activates a phosphatidylinositol-calcium second messenger system. The activation of LXA4R could result in an anti-inflammatory outcome counteracting the actions of proinflammatory signals such as LTB4 (leukotriene B4).
SIZE: 351 amino acids; 38964 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: Expressed abundantly in the lung and neutrophils. Also found in the spleen and testis.
SIMILARITY: SwissProt: P25090 ## Belongs to the G-protein coupled receptor 1 family.

Application

Radioligand binding assay and GTPγS binding.

Packaging

200 units

Quality

Kd for WKYMVm: 0.6 nM in a GTPγS binding assay

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 0.5 mg/mL in 1 mL packaging buffer, rapidly frozen, and stored at -80°C.

Storage and Stability

Maintain frozen at -70°C for up to 2 years. Do not freeze and thaw.

Analysis Note

ASSAY CONDITIONS: Membranes are permeabilized by addition of saponin to an equal concentration by mass, then mixed with [35S]-GTPγS (final concentration of 0.3 nM) in 20 mM HEPES, pH 7.4/100 mM NaCl/10 mM MgCl2/0.5 μM GDP in a nonbinding 96-well plate. Unlabeled WKYMVm is added to the final concentration indicated in Figure 1 (final volume 100 μL), and incubated for 30 min at 30°C. The binding reaction is transferred to an FB filter plate (Millipore MAHF B1H) pre-wetted with water, and washed 2 times (1 mL per well per wash) with cold 10 mM sodium phosphate, pH 7.4. The plate is dried and counted.

One vial contains enough membranes for at least 200 assays (units), where one unit is the amount of membrane that will yield greater than 1000 cpm specific WKYMVm stimulated [35S]-GTPγS binding when final concentration of 0.3nM [35S]-GTPγS was used.

Disclaimer

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.

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