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现在位置首页>实验室原料试剂耗材>蛋白检测>蛋白检测试剂盒>Human Adiponectin ELISA Kit
Human Adiponectin ELISA Kit
Human Adiponectin ELISA Kit
  • Human Adiponectin ELISA Kit

Human Adiponectin ELISA Kit

产品报价:4277元

更新时间:2012/7/19 9:18:09

地:美国

牌:assaypro

号:EA2500-1

厂商性质: 生产型,贸易型,服务型,

公司名称: 北京乐博生物科技有限公司

产品关键词:

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AssayMax Human Adiponectin ELISA Kit
Catalog No. EA2500-1
Introduction
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).
Principal of the Assay
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.
Caution and Warning
• This kit is for research use only.
• The kit should not be used beyond the expiration date.
• The Stop Solution is an acid solution.
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).
2
• 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).
Storage Condition
• Store kit at 2-80C or -200C upon arrival up to the expiration date.
• Opened MIX Diluent may be stored for up to 1 month at 2 - 80C. Store reconstituted reagents at -200C or below.
• Opened unused strip wells may return to the foil pouch with the desiccant pack, reseal along zip-seal. May be stored for up to 1 month in a vacuum desiccator.
Other Supplies Required
• Microplate reader capable of measuring absorbance at 450 nm.
• Pipettes (1-20 μl, 20-200 μl, 200-1000μl and multiple channel).
• Deionized or distilled reagent grade water.
Sample Collection and Storage
• Plasma: Collect plasma using one-tenth volume of 0.1 M sodium citrate as an anticoagulant. Centrifuge samples at 2000 x g for 10 minutes and assay. Dilute samples 1:500 into MIX Diluent. The undiluted samples can be stored at -200C or below for up to 3 months. Avoid repeated freeze-thaw cycles. (EDTA or Heparin can also be used as anticoagulant.)
• Serum: Samples should be collected into a serum separator tube. After clot formation, centrifuge samples at 2000 x g for 10 minutes. Remove serum and assay. Dilute samples 1:500 into MIX Diluent. The undiluted samples can be stored at -200C or below for up to 3 months. Avoid repeated freeze-thaw cycles.
• Cell Culture Supernatants: Centrifuge cell culture media at 2000 x g for 10 minutes to remove debris. Collect supernatants and assay. Dilute samples 1:10 into MIX Diluent. Store samples at -200C or below. Avoid repeated freeze-thaw cycles.
• Urine: Collect urine using sample pot. Centrifuge samples at 600 x g for 10 minutes and assay. Store samples at -200C or below for up to 3 months. Avoid repeated freeze-thaw cycles.
Reagent Preparation
• Freshly dilute all reagents and bring all reagents to room temperature before use. If crystals have formed in the concentrate, mix gently until the crystals have completely dissolved.
• MIX Diluent Concentrate (10x): Dilute the MIX Diluent Concentrate 1:10 with reagent grade water. Store for up to 1 month at 2 - 80C.
• Adiponectin Standard: Reconstitute the 800 ng of human adiponectin Standard with 4 ml of MIX Diluent to generate a standard stock of 200 ng/ml. Allow the standard to sit for 10 minutes with gentle agitation prior to making dilutions. Prepare duplicate or triplicate standard points by diluting stock (200 ng/ml) 1: 4 to produce a standard solution of 50 ng/ml. Prepare duplicate or triplicate standard points by serially diluting the Standard solution (50 ng/ml) 1:2 with equal volume of MIX Diluent to produce 25, 12.5, 6.25, 3.125, 1.56 and 0.78 ng/ml. MIX Diluent serves as the zero standard (0 ng/ml). Any remaining solution should be frozen at -200C.
, Serum, Urine and Cell Culture Supernatant samples)

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