As you all know that ELISA kit is a short name for enzyme-linked immunosorbent assays which are designed for the quantitative measurement of antigens or antibodies in a wide variety of samples. These samples can be serum, blood samples, or even urine samples. Nowadays scientists demand high-quality consistent data from these tests.
Hepcidin kit uses a quantitative sandwich ELISA technique and comes with highly specific antibodies already coated with a 96-well microtiter plate. In the sandwich ELISA technique, two antibodies have used that sandwich the antigen which is a pair of matched antibodies. An antibody that is captured is coated on a microplate and then the sample is added over it. Hepcidin antibody is reactive to humans, rats.
The protein which is of interest then combines and is paralyzed on the plate. A conjugated antibody is then added that combines with an epitope on the targeted protein. A substrate is then added which produces a signal that is proportional to the amount of antigen present in the sample. This technique is highly specific in nature.
It has its advantages like it is very sensitive in nature. It is compatible with complicated samples. But this technique takes time as compared to the other types of ELISA techniques and sometimes it is challenging to develop. When you have placed your samples on the microtiter plate, you need to incubate them with a highly specific biotin-conjugated primary antibody.
After cleaning the plate, add TMB (Tetramethylbenzidine) solution, into your solution. The amount of Hepcidin is directly proportional to the signal generated by the reaction. It means the kit assay gives you a quantitative measurement of the antibody present in your samples.