|Product Name:||Human IgG ELISA Kit
||IgG; Immunoglobulin G
|Sample Volume Required Per Well:
||serum, plasma and other biological fluids
||This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human IgG . Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human IgG and Avidin-Horseradish Peroxidase (HRP) conjugate is added to each micro plate well successively and then incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human IgG , biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color turns to yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human IgG . You can calculate the concentration of Human IgG in the samples by comparing the OD of the samples to the standard curve.
|Procedure:||1. Add 100μL standard or sample to each well. Incubate 90 mintues at 37℃
2. Add 100μL Biotinylated Detection Ab. Incubate 1 hour at 37℃
3. Aspirate and wash 3 times
4. Add 100μL HRP Conjugate. Incubate 30 minutes at 37℃
5. Aspirate and wash 5 times
6. Add 90μL Substrate Reagent. Incubate 15 minutes at 37℃
7. Add 50μL Stop Solution. Read at 450nm immediately.
8. Calculation of results