Human ELISA kit
A Human ELISA (Enzyme-Linked Immunosorbent Assay) kit is a laboratory tool used for the quantitative measurement of specific proteins or molecules in human samples, such as serum, plasma, urine, or cell lysates. ELISA kits are widely used in research, diagnostics, and drug discovery processes.
Here's how a typical Human ELISA kit works:
ELISA kits employ an antibody-antigen interaction principle. The kit contains pre-coated microplates with capture antibodies specific to the target protein of interest.
Human biological samples (such as serum or plasma) are collected from individuals and prepared according to the kit instructions. This may involve diluting the samples to ensure that they fall within the detection range of the assay.
The prepared samples, along with standards of known concentrations, are added to the wells of the microplate. These standards help to generate a standard curve for quantification.
During the first incubation step, the target protein in the sample binds to the capture antibody immobilized on the microplate surface.
After the incubation period, the microplate is washed to remove any unbound substances, such as nonspecific proteins or contaminants.
A detection antibody, conjugated to an enzyme such as horseradish peroxidase (HRP) or alkaline phosphatase (AP), is added to each well. This antibody recognizes a different epitope on the target protein and binds to it, forming a sandwich complex (capture antibody - target protein - detection antibody).
After another incubation period, the microplate is washed again to remove any unbound detection antibodies.
A substrate solution containing a chromogenic or fluorogenic substrate specific to the enzyme conjugated to the detection antibody is added to each well. The enzyme catalyzes a reaction with the substrate, resulting in the generation of a detectable signal (color change for chromogenic substrates, or fluorescence for fluorogenic substrates).
The intensity of the generated signal is directly proportional to the amount of target protein present in the sample. The signal is measured using a microplate reader, which quantifies the optical density (OD) of each well at the appropriate wavelength.
The OD values obtained from the standards are used to generate a standard curve, which is then used to interpolate the concentrations of the target protein in the samples.
Human ELISA kits are available for a wide range of analytes, including cytokines, growth factors, hormones, enzymes, and disease markers. They provide a sensitive, specific, and quantitative method for the detection and measurement of proteins in biological samples, making them valuable tools in biomedical research and clinical diagnostics.
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