PathHunter® C5AR1 Bioassay Kit (β-Arrestin Recruitment)

Background Information:

The PathHunter® C5AR1 Bioassay Kit (β-Arrestin Recruitment) is a robust, highly sensitive functional cell-based assay for quantitative potency testing and detection of neutralizing antibodies against biologic therapeutics targeting complement C5a receptor 1 (C5aR1/CD88). C5aR1 is a G protein-coupled receptor (GPCR) activated by the complement fragment C5a, a key mediator of innate immune and inflammatory responses. Activation of the C5a-C5aR1 pathway drives neutrophil activation, chemotaxis, and amplification of complement-mediated inflammation.
This bioassay kit uses EFC technology to measure C5AR1 activity by detecting the interaction of β-Arrestin with the activated GPCR using β-galactosidase (β-gal) enzyme fragment complementation. In this system, C5AR1 is fused in frame with the small β-gal fragment called ProLink™ (PK) and co-expressed in cells stably expressing a fusion protein of β-Arrestin-2 with the larger fragment of β-gal, enzyme acceptor (EA). Activation of C5AR1 triggers the recruitment of β-Arrestin-2 to the PK-tagged receptor and forces complementation of the two enzyme fragments of β-galactosidase. This results in the formation of an active β-gal enzyme leading to an increase in enzyme activity that can be measured using the chemiluminescent PathHunter Bioassay Detection Reagents.
Bioassay kits are a convenient, ready-to-use format which contains all materials needed to run the assay, including single-use vials of cryopreserved cells, cell plating media, control agonist, detection reagent, and assay plates.
Please reach out to Tech Support at DiscoverXSupportUS@discovery.eurofinsus.com for lot-specific CoA.

Target Class:

GPCR

Family:

Complement Peptide

Sub Family:

Class A

Protein Name:

C5aR

Protein Aliases:

C5a-R|C5aR|CD antigen CD88

Accession Number:

NM_001736

UniProt Number:

P21730

Gene Name:

C5AR1

Gene ID:

728

Gene Aliases:

C5A|C5AR|CD88

Target Species:

Human

Cell Background:

CHO-K1