Elisa Substrate Detection
Elisa Substrate Detection - Related Videos
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,...
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...
04:37
Detecting Neurodegenerative Disease-Associated Protein α-Synuclein Using ELISA
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2025
Source: Bétemps, D., et al., Detection of Disease-associated α-synuclein by Enhanced ELISA in the Brain of Transgenic Mice Overexpressing Human A53T Mutated α-synuclein. J. Vis. Exp. (2015).This video demonstrates the procedure to detect neurodegenerative disease-associated protein α-synuclein in brain tissue from transgenic mice using ELISA. The steps include tissue homogenization, centrifugation, and ELISA detection of α-synuclein protein to quantify its disease-associated forms.
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.
12:38
Titration ELISA as a Method to Determine the Dissociation Constant of Receptor Ligand Interaction
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Cited by 29 •
2018
A detailed protocol to perform a titration ELISA is described. Moreover, a novel algorithm is presented to evaluate titration ELISAs and to obtain a dissociation constant of binding of a soluble ligand to a microtiter plate-immobilized receptor.
FAQs
The enzyme-conjugated antibody serves as the reporting component after it binds the captured target. Adding substrate gives the attached enzyme a reactant it can convert into a detectable product. Because the reaction occurs where target-antibody binding has been retained, the resulting signal connects molecular recognition with an observable assay result.
These readouts differ in how the assay result is observed or recorded. Colored products provide a visible signal, whereas fluorescent and luminescent products produce signals through their respective optical readout modes. The underlying recognition step remains the same: an enzyme-conjugated antibody associates with the captured target, and substrate conversion generates the measurable output.
Signal intensity is meaningful only when assay conditions are defined consistently. Under those conditions, differences in product formation can be related to differences in the amount of analyte present. Without that controlled basis, signal strength cannot be interpreted reliably as a measure of target abundance. This principle distinguishes quantitative measurement from simple detection.
The enzyme-linked antibody connects molecular recognition to signal generation. Its antibody portion binds the target captured in the assay, while its enzyme portion acts on the added substrate. This separation of roles lets the assay convert an antibody-antigen interaction into a visible or instrumentally measurable product, making the target assessable even though the binding event itself is not directly visible.
A basic workflow begins with a target captured in the assay, followed by binding of an enzyme-conjugated antibody. After that association is established, the appropriate substrate is added, and the product signal is observed or measured. The resulting color, fluorescence, or luminescence provides the readout used to determine whether the target is present and, under defined conditions, how much is present.
The approach can examine proteins, antibodies, hormones, and pathogen markers in biological samples. In biology, these measurements support research, clinical testing, immunology, and diagnostic assay development. The same substrate-detection principle therefore serves investigations of biological molecules and practical assays designed to identify or quantify markers in samples.