High Throughput Elisa
High Throughput Elisa - 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,...
09:00
High Throughput Sequential ELISA for Validation of Biomarkers of Acute Graft-Versus-Host Disease
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Cited by 14 •
2012
High throughput validation of multiple candidate biomarkers can be performed by sequential ELISA in order to minimize freeze/thaw cycles and use of precious plasma samples. Here, we demonstrate how to sequentially perform ELISAs for six different validated plasma biomarkers1-3 of graft-versus-host disease (GVHD)4 on the same plasma sample.
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...
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.
09:44
High-throughput Screening for Chemical Modulators of Post-transcriptionally Regulated Genes
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2015
Here we describe a cell-based reporter gene assay as a valuable tool to screen chemical libraries for compounds modulating post-transcriptional control mechanisms exerted through 3’ UTR.
FAQs
The assay uses antibodies with complementary roles: one captures the target antigen, while another detects it through an enzyme-linked format. This arrangement connects target recognition to signal generation, allowing the amount of antigen in each biological sample to be estimated. Using these paired recognition steps supports measurements of proteins, hormones, cytokines, antibodies, and other biomarkers.
After the target is recognized, the linked enzyme reacts with a substrate to produce a colorimetric, fluorescent, or chemiluminescent signal. The measured signal is proportional to the target concentration, so differences between samples can be compared quantitatively. The selected readout therefore determines how the plate reader records the assay response rather than changing the antibody-based recognition process.
Miniaturized microplate formats accommodate many samples while reducing the scale of individual assay reactions. Automated liquid handling helps process those wells consistently, and standardized incubation and washing steps limit variation between samples or plates. Together, these features increase capacity and support reproducible results, which is especially important when biological studies require large-scale comparisons.
Consistency depends on controlling the repeated operations that connect antibody binding to signal measurement. Liquid transfer, incubation, washing, and plate reading must follow standardized conditions across the microplate. If these stages vary between wells or plates, the resulting signal may be less comparable. Reproducible handling is therefore central to reliable quantitative analysis in large sample sets.
A typical workflow coordinates sample handling in microplate wells, antibody-based capture and detection, standardized incubation, washing, substrate development, and plate reading. Liquid-handling automation manages transfers, while the reader measures the resulting colorimetric, fluorescent, or chemiluminescent output. This coordinated sequence lets laboratories process many biological samples efficiently without treating each sample as a separate manual assay.
The method is useful when investigators need quantitative information from many samples, such as during biomarker screening, drug development, diagnostics, or large-scale biological studies. Its supported targets include proteins, hormones, antibodies, cytokines, and other biomarkers. High sample capacity makes it suitable for comparing broad sample collections while maintaining a standardized antibody-based measurement approach.