DESCRIPTION

Adiponectin, also known as Acrp30, AdipoQ, and GBP28, is an adipocyte-secreted protein that circulates in high concentrations in the serum and acts to increase insulin sensitivity. Adiponectin modulates a number of metabolic processes, including glucose regulation and fatty acid oxidation. The serum concentration of adiponectin decreases in obese patients, opposite to other adipocytokines. In recent studies, serum adiponectin level progressively decreased in patients according to the severity of obesity, insulin resistance, diabetes mellitus and cardiovascular diseases. Low serum adiponectin level may be a predictive factor for type 2 diabetes mellitus (T2DM) and other cardiovascular diseases.

PRINCIPLE OF THE ASSAY

Sandwich ELISA structure (Detection antibody labeled with HRP) A capture antibody is pre-coated onto the bottom of wells which binds to analyte of interest. A detection antibody labeled with HRP also binds to the analyte. 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	538.7	16.6	3.1
2	20	119.1	2.5	2.1
3	20	29.9	0.7	2.5

Inter-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	24	522.3	23.2	4.4
2	24	115.2	2.5	2.1
3	24	27.8	0.8	3.0

RECOVERY

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

Sample Type		Average% of Expected	Range (%)
Mouse serum	1:102,400	86	81-90
	1:204,800	87	84-89
Tissue homogenate	1:8,000	83	80-89
	1:16,000	87	81-92
Cell culture supernatant	1:64	83	77-90
	1:128	92	82-102

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 samples were initially diluted 1:12,800. The tissue homogenate samples were initially diluted 1:1,000. Cell culture supernatant was initially diluted 1:8.)

		Mouse serum	Tissue homogenate	Cell culturesupernatant
1:2	Average% of Expected	100	100	100
	Range (%)	-	-	-
1:4	Average% of Expected	106	100	101
	Range (%)	104-108	100-101	98-104
1:8	Average% of Expected	108	106	104
	Range (%)	105-111	104-108	100-107
1:16	Average% of Expected	112	113	103
	Range (%)	108-116	110-116	99-106

CALIBRATION

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

Ordering & shipping

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