DESCRIPTION

Interleukin-6 (IL-6) is a pleiotropic cytokine produced by a variety of cells during infection, trauma, and immunological challenge. IL-6 acts via a receptor complex consisting of two distinct membrane-bound glycoproteins, an 80-kDa IL-6-binding subunit (IL-6R, CD126, gp80) and a 130-kDa signal-transducing element (gp130, CD130). After binding of IL-6 to membranebound IL-6R, the complex of IL-6 and IL-6R associates with gp130, thus activating the receptor. Expression of gp130 is found in almost all organs while expression of IL-6R is predominantly confined to hepatocytes and leukocyte subpopulations (monocytes, neutrophils, T cells, and B cells). Soluble form of IL-6R has been found in human serum and urine.

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	508.9	12.9	2.5
2	8	85.5	1.2	1.4
3	8	60.8	1.6	2.6

Inter-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	16	517.8	16.5	3.2
2	16	129.4	5.5	4.2
3	16	60.0	1.8	3.0

RECOVERY

The recovery of human CD126/IL-6R alpha spiked to three different levels throughout the range of the assay was evaluated.

Sample Type		Average% of Expected	Range (%)
Human plasma	1:1,280	85	73-96
	1:2,560	96	88-103
Cell culture supernatant	1:8	98	94-104
	1:16	103	91-114
Urine	1:128	100	94-110
	1:256	101	93-108

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 human plasma was initially diluted 1:160. The urine was initially diluted 1:16.)

		Human plasma	Cell culture supernatant	Urine
1:2	Average% of Expected	100	100	100
	Range (%)	-	-	-
1:4	Average% of Expected	111	106	82
	Range (%)	102-119	100-113	80-84
1:8	Average% of Expected	119	94	101
	Range (%)	114-123	83-105	99-103
1:16	Average% of Expected	127	85	107
	Range (%)	125-130	81-90	104-110

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human Human CD126/IL-6R alpha One-Step ELISA Kit produced at Sunlong Medical™.

Ordering & shipping

• Minimum order: 1 kit
• Lead time: varies with order quantity and specifications. Please consult your local distributor or our sales team for confirmation
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• Shipping: FedEx / DHL / UPS express; cold-chain packing available on request
• Price: USD, per kit (48T / 96T options available)