Description |
The principle of the double antibody sandwich ELISA is represented in Figure 1. In this assay the IgG present in samples reacts with the anti-IgG antibodies which have been adsorbed to the surface of polystyrene microtitre wells. After the removal of unbound proteins by washing, anti-IgG antibodies conjugated with horseradish peroxidase (HRP), are added. These enzyme-labeled antibodies form complexes with the previously bound IgG. Following another washing step, the enzyme bound to the immunosorbent is assayed by the addition of a chromo¬genic substrate, 3, 3’, 5, 5’-tetramethylbenzidine (TMB). The quantity of bound enzyme varies directly with the concentration of IgG in the sample tested, thus, the absorbance, at 450 nm, is a measure of the concentration of IgG in the test sample. The quantity of IgG in the test sample can be interpolated from the standard curve constructed from the standards, and corrected for sample dilution. |
Reconstitution and storage |
1. DILUENTThe 5X Diluent Concentrate is stable until the expira-tion date. The 1X working solution is stable for at least one week from the date of preparation. Both solutions should be stored at 4-8C.2. WASH SOLUTIONThe 20X Wash Solution Concentrate is stable until the expira¬tion date. The 1X working solution is stable for at least one week from the date of preparation. Both solutions can be stored at room temperature (16 25C) or at 4-8C.3. ENZYME-ANTIBODY CONJUGATEUndiluted horseradish peroxidase anti-IgG conjugate should be stored at 4-8C and diluted immediately prior to use. The working conjugate solution is stable for up to 8 hours.4. CHROMOGEN-SUBSTRATE SOLUTIONThe Substrate Solution should be stored at 4-8C and is stable until the expiration date.5. STOP SOLUTIONThe Stop Solution should be stored at 4-8C and is stable until the expiration date.6. ANTI-HUMAN IgG ELISA MICRO PLATEAnti-Human IgG coated wells are stable until the expiration date, and should be stored at 4-8C in sealed foil pouch with desiccant pack.7. HUMAN IgG CALIBRATORLong Term Storage: Upon receipt, aliquot the calibrator and store them frozen. They will be stable until expiration date. Short Term Storage: the calibrator is stable for up to 14 days at 4C. The working standard solutions should be prepared immediately prior to use and are stable for up to 8 hours. |