Intended use
This immunoassay kit allows for the in vitro quantitative determination of Human Isoform 1 of Vitamin D-binding protein，DBP-I1, concentrations in serum, tissue homogenates and other biological fluids.
Test principle
The microtiter plate provided in this kit has been pre-coated with an antibody specific to DBP-I1. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated polyclonal antibody preparation specific for DBP-I1 and Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then a TMB substrate solution is added to each well. Only those wells that contain DBP-I1, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of DBP-I1 in the samples is then determined by comparing the O.D. of the samples to the standard curve.
Materials and components
Reagent Quantity
Assay plate 1
Standard 2
Sample Diluent 1 × 20ml
Assay Diluent A 1 × 10ml
Assay Diluent B 1 × 10ml
Detection Reagent A 1 × 120μl
Detection Reagent B 1 × 120μl
Wash Buffer(25 x concentrate) 1 × 30ml
Substrate 1 × 10ml
Stop Solution 1 × 10ml
Plate sealer for 96 wells 5
Instruction 1
