DESCRIPTION

Resistin, a cysteine-rich hormone secreted primarily by rodent fat cells, was postulated to be implicated in obesity, insulin resistance and diabetes. Resistin is also secreted by macrophages in humans, promoting inflammation (PMID: 20148674). Resistin modulates the synthesis and secretion of key pro-inflammatory cytokines such as TNF-α, IL-6 and IL12 in macrophages via a nuclear factor-kB (NF-kB)-dependent pathway. Given the connection between inflammation and cancer, resistin was soon considered for its role in cancer. There were observations supporting its possible use as a diagnostic marker, expression levels of which also seemed to differ in patients with varying clinical stages (PMID: 30317640). Resistin has been also linked to other pathologies such as atherosclerosis (PMID: 15710760).

PRINCIPLE OF THE ASSAY

Sandwich ELISA structure (Detection antibody labeled with biotin) A capture antibody is pre-coated onto the bottom of wells which binds to analyte of interest. A detection antibody labeled with biotin also binds to the analyte. Streptavidin-HRP binds to the biotin. TMB acts as the HRP substrate and the solution color will change from colorless to blue. A stop solution containing sulfuric acid turns 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 20 times on one plate to assess intra-assay precision. Inter-assay Precision (Precision between assays) Three samples of known concentration were tested in 24 separate assays to assess inter-assay precision.

Intra-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	20	977.2	35.8	3.7
2	20	216.5	10.9	5.0
3	20	51.5	3.7	7.2

Inter-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	24	961.4	47.4	4.9
2	24	224.2	10.5	4.7
3	24	52.3	3.3	6.2

RECOVERY

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

Sample Type		Average % of Expected	Range (%)
Human plasma	1:8	87	81-92
	1:16	90	79-103
Cell culture supernatant	1:2	84	77-89
	1:4	87	77-96

LINEARITY

To assess the linearity of the assay, human plasma samples were diluted with the appropriate Sample Diluent to produce samples with values within the dynamic range of the assay. Cell culture supernatant was spiked with high concentrations of Resistin with the appropriate Sample Diluent to produce samples with values within the dynamic range of the assay.

		Human plasma	Cell culture supernatant
1:2	Average% of Expected	100	93
	Range (%)	-	88-97
1:4	Average% of Expected	113	99
	Range (%)	106-121	91-105
1:8	Average% of Expected	109	100
	Range (%)	103-114	94-105
1:16	Average% of Expected	98	102
	Range (%)	96-100	95-109

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human Resistin produced at Sunlong Medical™.

Ordering & shipping

• Minimum order: 1 kit
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• Price: USD, per kit (48T / 96T options available)