DESCRIPTION

Chemerin, also named as TIG2 and RARRES2, is adipocyte-secreted protein (adipokine) that regulates adipogenesis, metabolism and inflammation through activation of the chemokine-like receptor 1 (CMKLR1). Its other ligands include G protein-coupled receptor 1 (GPR1) and chemokine receptor-like 2 (CCRL2). It positively regulates adipocyte differentiation, modulates the expression of adipocyte genes involved in lipid and glucose metabolism and might play a role in angiogenesis, a process essential for the expansion of white adipose tissue. Chemerin have both proand anti-inflammatory properties depending on the modality of enzymatic cleavage by different classes of proteases.

PRINCIPLE OF THE ASSAY

An anti-tag antibody is pre-coated onto the bottom of wells. After adding antigen or samples, Tagged-Capture antibody and HRP-conjugated detection antibody, a sandwich complex is formed in the solution. TMB acts as a HRP substrate, and the solution color will change from colorless to blue. A stop solution containing sulfuric acid turns the solution yellow. The color intensity is proportional to the quantity of bound protein, which is measurable at 450 nm with the correction wavelength set at 630 nm.

PRECISION

Intra-assay Precision (Precision within an assay) Three samples of known concentration were tested 8 times on one plate to assess intra-assay precision. Inter-assay Precision (Precision between assays) Three samples of known concentration were tested in 16 separate assays to assess inter-assay precision.

Intra-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	8	263.9	6.4	2.4
2	8	58.1	2.1	3.6
3	8	26.6	0.6	2.3

Inter-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	16	257.3	8.8	3.4
2	16	57.9	2.1	3.6
3	16	27.1	0.9	3.3

RECOVERY

The recovery of mouse Chemerin spiked to three different levels throughout the range of the assay in various matrices was evaluated.

Sample Type		Average% of Expected	Range (%)
Mouse serum	1:1,600	106	87-120
	1:3,200	105	104-106
Cell culture supernatant	1:8	109	104-117
	1:16	98	93-104

LINEARITY

To assess the linearity of the assay, samples were diluted with the appropriate Sample Diluent to produce samples with values within the dynamic range of the assay. (The mouse serum was initially diluted 1:200.)

		Mouse serum	Cell culture supernatant
1:2	Average% of Expected	100	100
	Range (%)	-	-
1:4	Average% of Expected	100	106
	Range (%)	99-100	104-108
1:8	Average% of Expected	100	112
	Range (%)	96-103	111-113
1:16	Average% of Expected	102	110
	Range (%)	97-107	102-117

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant mouse Chemerin produced at Sunlong Medical™.