Adiponectin, BioAssay(TM) ELISA Kit (Rat)

Adiponectin(ADPN) is a hormone secreted by adipocytes that regulates energy homeostasis and glucose and lipid metabolism. Adiponectin is a new member of the family of soluble defense collagens, in hematopoiesis and immune responses. It is an important negative regulator in hematopoiesis and immune systems and raise the possibility that it may be involved in ending inflammatory responses through its inhibitory functions. Adiponectin is mapped to 3q27 and can protect the organism from systemic inflammation by promoting the clearance of early apoptotic cells by macrophages through a receptor-dependent pathway involving calreticulin. The Adiponectin, BioAssay(TM) ELISA Kit is a sandwich ELISA for quantitative detection of rat Adiponectin in cell culture supernates, serum, plasma (heparin, EDTA) and urine. , Detection Range:, 156-10,000pg/ml, Sensitivity: <10pg/ml, Specificity: Natural and recombinant rat Adiponectin, Crossreactivity: There is no detectable cross-reactivity with other relevant proteins, Assay Principle: The Adiponectin BioAssay(TM) ELISA Kit is a solid phase immunoassay specially designed to measure rat Adipoq with a 96-well strip plate that is pre-coated with antibody specific for Adipoq. The detection antibody is a biotinylated antibody specific for Adipoq. The capture antibody is monoclonal antibody from mouse, the detection antibody is polyclonal antibody from goat. The kit contains recombinant rat Adipoq with immunogen (Expression system for standard: E. coli, Immunogen sequence: I19-N244). The kit is analytically validated with ready to use reagents. To measure rat Adipoq, add standards and samples to the wells, then add the biotinylated detection antibody. Wash the wells with PBS or TBS buffer, and add Avidin-Biotin-Peroxidase Complex (ABC-HRP). Wash away the unbounded ABC-HRP with PBS or TBS buffer and add TMB. TMB, is substrate to HRP and will be catalyzed to produce a blue color product, which changes into yellow after adding acidic stop solution. The density, of the yellow product is linearly propotional to rat Adipoq in the sample. Read the density of the yellow product in each well using a plate reader,, and benchmark the sample wells' readings against the standard curve to determine the concentration of rat Adipoq in the sample. Kit Components: 352697A: Microtiter Strips, 1x96 wells, 352697B: Adiponectin Standard, 2x20ng, 352697C: Anti-rat Adiponectin (Biotin), 1x130ul (dilute 1:100), 143696: Avidin-Biotin-Peroxidase Complex (ABC), 1x130ul (1:100 dilution), 143697: Sample Diluent Buffer, 1x30ml, 143698: Antibody Diluent Buffer, 1x12ml, 143699: ABC Diluent Buffer, 1x12ml, 143700: TMB Color Developing Agent, 1x10ml, 143701: TMB Stop Solution, 2N H2SO4, 1x10ml, Storage and Stability: Store all components at 4°C. Stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Assay Summary: 1. Add 100ul of samples and standards and incubate the plate at 37°C for 90 minutes or at RT for 2 hours. Do not wash., 2. Add 100ul biotinylated antibody and incubate the plate at 37°C for 60 minutes or at RT for 90 minutes. Wash plate 3 times with 0.01M PBS or TBS., 3. Add 100ul of ABC working solution and incubate the plate at 37°C for 30 minutes or at RT for 40 minutes. Wash plate 5 times with 0.01M PBS or TBS., 4. Add 90ul of TMB color developing agent and incubate the plate at 37°C in dark for 25-30 minutes or at RT for 30 minutes., 5. Add 100ul TMB Stop Solution and read.