The Human Estradiol (E2) ELISA Kit is a kit for the quantitative detection of estradiol concentrations in human serum or plasma. The principle is based on an enzyme-linked immunosorbent assay (ELISA), which converts the target hormone into a measurable signal through the binding reaction of a specific antibody to the target hormone. The principles and procedures of the Human Estradiol (E2) ELISA Kit will be described in detail in this article.
1. The principle of human estradiol (E2) ELISA kit.
The principle of the Human Estradiol (E2) ELISA Kit is based on an enzyme-linked immunosorbent assay (ELISA). This method is a sensitive, specific, and reproducible immunological analysis method. The kit uses the principle of competitive inhibition to bind a specific antibody to a target hormone (estradiol) to form an antigen-antibody complex. The complex binds to an antibody immobilized on the plate and inhibits the binding of the antibody to the label. The higher the concentration of the target hormone in the sample, the less antibody binds to the plate and the darker the color of the remaining labeled antibody reacts with the substrate. By measuring the optical density value, the concentration of estradiol in the sample can be calculated.
2. Procedure for human estradiol (E2) ELISA kit.
1.Reagent preparation.
Before use, take the kit out of the refrigerator, place it at room temperature, open the kit, and add all the reagents in the kit as required. Gently shake the plate to mix all the reagents thoroughly.
2.Dispensing.
The standard and the sample to be tested were added to the corresponding microwells, and the blank wells were added with an equal volume of washing solution. When dispensing, care should be taken not to add the sample to the wall of the well, so as not to affect the test results.
3.Incubation.
Place the plate in a 37 incubator and incubate for 30 min. During the incubation process, the plate should be kept horizontally to avoid spillage of liquid caused by tilting or inversion.
4.Wash.
Remove the microplate from the incubator and rinse each well 2-3 times with wash solution, making sure to rinse the liquid from the wells. After washing, gently absorb the remaining water with a paper towel.
5.Color development.
Add the substrate solution to each well and gently shake the plate to mix the solution thoroughly. Place the plate in a 37 incubator for 15-20 min to develop color. During color development, direct light should be avoided.
6.Terminate the reaction.
Add the stop solution to each well and immediately blot off the excess liquid with a paper towel. In this case, air bubbles should be avoided to affect the test results.
7.Detect.
The optical density value of each well was read at a wavelength of 450 nm with a microplate reader and the data was recorded. A standard curve was made according to the optical density value of the standard, and the corresponding concentration value was found on the standard curve according to the optical density value of the sample to be measured.