Porcine Mannose Receptor C1-like Protein 1 (MRC1) ELISA Kit Procedure.
1. Experimental principle.
This kit uses indirect enzyme-linked immunoassay to determine the level of porcine mannose receptor C1-like protein 1 (MRC1) in porcine serum, plasma, cell culture supernatant or other related biological samples. The plate was coated with purified porcine mannose receptor C1-like protein 1 (MRC1) antibody, and then the sample to be tested and the enzyme-labeled porcine mannose receptor C1-like protein 1 (MRC1) antibody were added, incubated and thoroughly washed, and the substrate was added for color development. Finally, the optical density was measured by microplate reader at 450nm, and the absorbance value of the sample was negatively correlated with the concentration of porcine mannose receptor C1-like protein 1 (MRC1), and the concentration of porcine mannose receptor C1-like protein 1 (MRC1) in the sample was calculated by standard curve.
2. Steps.
1.Reagent preparation: Pretreat and store all reagents as directed on the label.
2.Loading: Add the sample to be tested and the standard to the corresponding microplate, and add the appropriate amount of diluent.
3.Incubation: After sealing the microplate, incubate at 37 for 30 min.
4.Washing: Wash the microplate thoroughly to remove unbound proteins and other impurities.
5.Enzyme label addition: Enzyme-labeled antibodies were added to the microplate, sealed, and incubated for 30 minutes.
6.Washing: Wash the microplate thoroughly again to remove unbound enzyme labels.
7.Chromogenesis: Substrate solution is added and incubated for 15 min at room temperature.
8.Stop the reaction: Add the stop solution to stop the color development reaction.
9.Detection: The optical density of each well is measured on a microplate reader and the data is recorded.
10.Data analysis: The concentration of porcine mannose receptor C1-like protein 1 (MRC1) in the sample to be tested was calculated according to the standard curve.
3. Precautions.
1.In order to ensure the accuracy of the test results, the operation procedures and precautions should be strictly followed during the experiment.
2.Before using the reagent, its appearance and expiration date should be carefully checked to ensure that the quality of the reagent meets the requirements.
3.The microplate should be kept sealed throughout the experiment to avoid contamination and evaporation.
4.Make sure that each well is adequately washed during the wash to remove unbound proteins and other impurities.
5.In the process of color reaction, the temperature and time should be controlled to ensure the accuracy of the reaction.
6.Optical density measurements should be performed at a set wavelength to ensure the accuracy and reliability of the data.