Indirect Elisa
Indirect Elisa - Related Videos
14:22
ELISA Assays: Indirect, Sandwich, and Competitive
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2023
Source: Whitney Swanson1,2, Frances V. Sjaastad2,3, and Thomas S. Griffith1,2,3,4 1 Department of Urology, University of Minnesota, Minneapolis, MN 55455 2 Center for Immunology, University of Minnesota, Minneapolis, MN 55455 3 Microbiology, Immunology, and Cancer Biology Graduate Program, University of Minnesota, Minneapolis, MN 55455 4 Masonic Cancer Center, University of Minnesota, Minneapolis, MN 55455 Enzyme-linked immunosorbent assay (ELISA) is frequently used to measure the presence...
10:06
The ELISA Method
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2023
An enzyme-linked immunosorbent assay (ELISA) is typically performed to detect the presence and/or amount of a target protein of interest within an experimental sample. Detection of the target protein is made possible by antibodies, which make the ELISA an immunoassay. Through a series of incubation and washing steps, these antibodies, which are frequently linked, or conjugated, to an enzyme, will detect protein coating the bottom of a well on a microtiter plate. When exposed to a substrate,...
01:30
Indirect Costs
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2026
Indirect costs are costs that cannot be directly traced to a single product, service, or cost object. Instead, they support multiple products or activities simultaneously and must be allocated using a systematic and reasonable method. Cost allocation is the process of distributing these shared costs among cost objects so that each product bears a fair share of the total overhead. Accurate allocation of indirect costs is essential for determining product costs, setting prices, evaluating...
03:55
ELISA-Based Method to Analyze Interactions between Bait and Prey Proteins
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2025
This video describes an ELISA-based technique that detects protein-protein interactions between two proteins by measuring the binding of the second protein to the immobilized first protein. The bound second protein is detected using specific antibodies labeled with an enzyme, such as horseradish peroxidase.
03:35
ELISA-Based Reporter Assay to Study the Receptor-Ligand Interaction in BW Cells
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2025
This video demonstrates the reporter assay-based detection of the receptor-ligand interaction. The successful receptor-ligand interaction activates the downstream signaling pathway, causing the generation and release of cytokines as secretory reporter molecules in the solution, which is confirmed using ELISA.
FAQs
The immobilized antigen provides the recognition target for antibodies in the biological sample. Only antibodies with binding specificity for that antigen can attach during the sample-incubation step, while the enzyme-linked secondary antibody identifies those sample antibodies after binding. This arrangement allows the assay to assess antibody recognition of a particular antigen rather than antibodies generally.
The substrate converts the enzyme activity of the bound secondary antibody into a measurable color change. Because the signal is proportional to the amount of antibody present, stronger color indicates more target antibody in the sample, whereas weaker color indicates less. The color therefore provides an estimate of antibody abundance for biological sample analysis.
The secondary antibody connects antibody recognition in the sample to a measurable signal. It attaches to sample antibodies that have already bound the immobilized antigen and carries the enzyme responsible for the substrate reaction. Without this linked component, antigen binding would not be translated into the color-based readout used to estimate antibody levels.
The known antigen immobilized on the microplate determines which antibody-binding response the assay examines. A sample may contain many antibodies, but the measured signal reflects those able to bind that selected antigen. Consequently, changing the antigen changes the biological question, allowing analysis of antibody specificity toward different targets within immune-response studies.
The procedure begins by immobilizing a known antigen on a microplate. A biological sample is then added so antibodies with matching specificity can bind, followed by an enzyme-linked secondary antibody that attaches to those bound antibodies. Finally, substrate is added, and the resulting color change is measured to estimate the antibody amount.
Researchers can apply Indirect ELISA when they need to detect or estimate antibodies in biological samples. Its uses include serological testing, investigating exposure to pathogens, evaluating antibody responses during vaccine research, and examining antibody specificity. These applications make the format useful for connecting measurable antibody signals with immune responses and disease-related biomarkers.
By measuring the color signal generated after antigen-specific binding, the assay provides an estimate of the antibody response associated with a sample. In biology, that information can support studies of immune responses, disease-related biomarkers, pathogen exposure, vaccine-related antibody production, and the specificity of antibodies directed toward a known antigen.