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Array-Based ELISAs for High-Throughput Analysis of Human Cytokines
Open accessAbstract
In this report, we describe the development of a mini-array system suitable for high-throughput quantification of proteins. This mini-array is a multiplexed, sandwich-type ELISA that measures the concentration of seven different human cytokines--TNF-alpha, IFN alpha, IFN gamma, IL-1 alpha, IL-1 beta, IL-6, and IL-10--from a single sample in each well of a 96-well plate. The mini-array is produced by spotting monoclonal antibodies (mAbs) in a 3 x 3 pattern in the bottom of the wells of 96-well polystyrene plates. Cytokines that are captured by the arrayed mAbs are detected by using biotinylated mAbs, followed by the addition of a streptavidin-horseradish peroxidase (HRP) conjugate and a chemiluminescent substrate. The light produced from the HRP-catalyzed oxidation of the substrate is measured at each spot in the array by imaging the entire plate with a commercially available CCD camera. Here, we demonstrate that these 96-well-plate format mini-arrays have performance characteristics that make them suitable for the high-throughput screening of anti-inflammatory compounds.
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Cited by
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Metadata
- Authors
- Moody, M. D., Arsdell, S. W. V., Murphy, K. P., Orencole, S. F., & Burns, C.
- Venue
- BioTechniques
- Publisher
- Informa UK Limited
- Published
- 2001-07
- Volume
- 31
- Issue
- 1
- Pages
- 186-194
- ISSN
- 0736-6205 (ISSN-L)
- 1940-9818
- OpenAlex
- W1543099628
- Cited by
- 187
- References
- 0
- FWCI
- 6.62
- Type
- journal article
- Language
- en
- Open access
- gold
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- Moody, M. D., Arsdell, S. W. V., Murphy, K. P., Orencole, S. F., & Burns, C. (2001). Array-Based ELISAs for High-Throughput Analysis of Human Cytokines. BioTechniques. https://doi.org/10.2144/01311dd03
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