The Takara EIA Kit is a complete system for measuring specific molecules present in biological samples such as plasma, serum, urine and cultured cell extracts. The majority of these kits are based on a sandwich-type enzyme immunoassay (EIA), which utilizes a pair of monoclonal antibodies, each recognizing a different epitope on the antigen molecule. Quantitative measurement of antigen binding is mediated by enzymatic color formation in 96-well microtiter plates.
One of the antibodies in the pair is supplied pre-labeled with peroxidase (POD), and the other clone is precoated onto the microtiter plate. Specimens (or standard solutions) are incubated in the coated microtiter plate wells, and the antigen is captured onto the plate. After washing the plate, the labeled antibody is added to the wells and another incubation step is performed, tagging the bound antigen with POD-labeled antibody.
Reaction between the plate-bound POD and chromogenic substrate (H2O2, and TMBZ) generates production of colored reaction product. The quantity of reaction product can be measured by a plate reader and is proportional to the amount of antigen present in the specimens or standards.
The incubation and detection steps on pre-coated microtiter plates require approximately 3 hours.
Total Assay Capacity
Assay Capacity for Test Sample
If all assays (including standards) are run in duplicate, 48 reactions are possible.
Test Sample Type
These assays can generally be conducted on human serum, plasma, or urine; culture supernatants; or cell extracts. Recommended sample types may vary by kit.
Specimen Volume Required
If each test sample is run in duplicate, approximately 210 µL (i.e., 100 µL per assay plus 10 µL for each sample transfer) is required. One-step sandwich EIAs require
Since conditions may vary from assay to assay, a standard curve must be established for every run. Refer to user manual for more information if culture supernatants are to be measured.
Store at 2 to 8°C