High Sensitive Adiponectin ELISA Assay Kit

$575.00

The Eagle Biosciences High Sensitive Adiponectin ELISA Assay Kit is intended for the quantification of Human Adiponectin in serum, plasma, cell culture supernatants. The Eagle Biosciences Adiponectin ELISA Assay Kit is for research use only and should not be used for diagnostic procedures.

SKU: ARG80131 Categories: , ,

High Sensitive Adiponectin ELISA Assay Kit

For Research Use Only

Size: 1 x 96 wells
Sensitivity: 0.08 ng/ml
Dynamic Range: 0.156-10 ng/ml
Incubation Time: 4 hours
Sample Type: Serum, Plasma, Cell Culture Supernatants
Sample Size: 100 µl

Additional Information

Assay Background

This gene is expressed in adipose tissue exclusively. It encodes a protein with similarity to collagens X and VIII and complement factor C1q. The encoded protein circulates in the plasma and is involved with metabolic and hormonal processes. Mutations in this gene are associated with adiponectin deficiency. Multiple alternatively spliced variants, encoding the same protein, have been identified.

Assay Principle

The Eagle Biosciences High Sensitive Adiponectin ELISA Assay Kit employs the quantitative sandwich enzyme immunoassay technique. An antibody specific for Adiponectin has been pre-coated onto a microtiter plate. Standards or samples are pipetted into the wells and any Adiponectin present is bound by the immobilized antibody. After washing away any unbound substances, a biotin-conjugated antibody specific for Adiponectin is added to each well and incubate. Following a washing to remove unbound substances, streptavidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After washing away any unbound antibody-enzyme reagent, a substrate solution (TMB) is added to the wells and color develops in proportion to the amount of Adiponectin bound in the initial step. The color development is stopped by the addition of acid and the intensity of the color is measured at a wavelength of 450nm ±2nm.The concentration of Adiponectin in the sample is then determined by comparing the O.D of samples to the standard curve.

Assay Procedure

  1. Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal it.
  2. Add 100 μl of standards, samples and zero controls (standard diluent buffer) into wells. Incubate for 1.5 h at 36 °C.
  3. Aspirate each well and wash, repeating the process four times for a total five washes. Wash by filling each well with 1× Wash Buffer (350 μl) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating, decanting or blotting against clean paper towels.
  4. Add 100 μl 1X Antibody conjugate into each well. Cover wells and incubate for 1 hour at 36°C.
  5. Aspirate each well and wash as step 3.
  6. Add 100 μl of 1X HRP-Streptavidin solution to each well. Cover wells and incubate for 30 minutes at 36°C.
  7. Aspirate each well and wash as step 3.
  8. Add 100 μl of TMB Reagent to each well. Incubate for 15 minutes at 36°C in dark.
  9. Add 100 μl of Stop Solution to each well. The color of the solution should change from blue to yellow.
  10. Read the OD with a microplate reader at 450nm immediately.

Typical Standard Curve

Manual

Product Manual