The Adiponectin, BioAssay™ ELISA Kit is a sandwich ELISA for quantitative detection of rat Adiponectin in cell culture supernates, serum, plasma (heparin, EDTA) and urine.

156-10,000pg/ml

<10pg/ml

Natural and recombinant rat Adiponectin

There is no detectable cross-reactivity with other relevant proteins

The Adiponectin BioAssay™ 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.

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

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.

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.