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Specificity |
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This assay recognizes both natural and recombinant human adiponectin. It can detect both globular domain and full-length adiponectin.
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Protocol |
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• The minimum detectable dose of adiponectin is typically 0.5 ng/ml.
• Intra-assay and inter-assay coefficients of variation were 4.2 % and 7.3 % respectively.
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Storage |
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Store kit at 2-80C or -200C upon arrival up to the expiration date.
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References |
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(1) Tsao, T.S. et al. (2002) EJP 440:213-221
(2) Adamczak, M. et al. AJH 16:72-75
(3) Matsubara, M. et al. (2003) Eur J Endocrinol. 148(6): 657-62
(4) Kojima, S. et al. (2003) Heart 89(6): 667
(5) Zoccali, C. et al. (2003) Kidney Int Suppl. 84: S98-102
(6) Pannacciulli, N. et al. (2003) J Clin Endocrinol Metab. 88(4): 174
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Contents/Specifications |
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The AssayMax Human Adiponectin ELISA Kit is designed for detection of adiponectin in human urine, plasma, serum and cell culture supernatants. This assay employs a quantitative sandwich enzyme immunoassay technique that measures adiponectin in less than 4 hours. A polyclonal antibody specific for adiponectin has been pre-coated onto a microplate. Adiponectin in standards and samples is sandwiched by the immobilized antibody and biotinylated polyclonal antibody specific for adiponectin, which is recognized by a streptavidin-peroxidase conjugate. All unbound material is then washed away and a peroxidase enzyme substrate is added. The color development is stopped and the intensity of the color is measured.
Reagents
• Adiponectin Microplate: A 96-well polystyrene microplate (12 strips of 8 wells) coated with a polyclonal antibody against human adiponectin.
• Sealing Tapes: Each kit contains 3 pre-cut, pressure-sensitive sealing tapes, which can be cut to fit the format of the individual assay.
• Adiponectin Standard: Human adiponectin in a buffered protein base (800 ng, lyophilized).
• Biotinylated Adiponectin Antibody (100x): A 100-fold concentrated biotinylated polyclonal antibody against adiponectin (80 μl).
• MIX Diluent Concentrate (10x): A 10-fold concentrated buffered protein base (30 ml).
• Wash Buffer Concentrate (20x): A 20-fold concentrated buffered surfactant (30 ml, 2 botlles).
• Streptavidin-Peroxidase Conjugate (SP Conjugate): A 100-fold concentrate (80 μl).
• Chromogen Substrate: A ready-to-use stabilized peroxidase chromogen substrate tetramethylbenzidine (8 ml).
• Stop Solution: A 0.5 N hydrochloric acid to stop the chromogen substrate reaction (12 ml).
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Notes |
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Adiponectin, also known as Adipocyte Complement-Related Protein of 30kDa (ACRP30), is a secreted serum protein expressed exclusively in differentiated adipocytes. Studies indicated that decreased plasma adiponectin concentration is associated with obesity, insulin resistance (1), essential hypertension (2), inflammation and atherosclerosis (3), and acute myocardial infarction (4). On the other hand, increased adiponectin level leads to nephrotic syndrome (5, 6).
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