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Array-based ELISAs for high-throughput analysis of human cytokines.
- Source :
-
BioTechniques [Biotechniques] 2001 Jul; Vol. 31 (1), pp. 186-90, 192-4. - Publication Year :
- 2001
-
Abstract
- 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.
- Subjects :
- Anti-Inflammatory Agents pharmacology
Antibodies, Monoclonal
Cell Line
Cytokines immunology
Humans
Interferon-alpha analysis
Interferon-alpha immunology
Interferon-gamma analysis
Interferon-gamma immunology
Interleukin-1 analysis
Interleukin-1 immunology
Interleukin-10 analysis
Interleukin-10 immunology
Interleukin-6 analysis
Interleukin-6 immunology
Monocytes cytology
Monocytes drug effects
Monocytes immunology
Sensitivity and Specificity
Tumor Necrosis Factor-alpha analysis
Tumor Necrosis Factor-alpha immunology
Cytokines analysis
Enzyme-Linked Immunosorbent Assay methods
Subjects
Details
- Language :
- English
- ISSN :
- 0736-6205
- Volume :
- 31
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- BioTechniques
- Publication Type :
- Academic Journal
- Accession number :
- 11464511
- Full Text :
- https://doi.org/10.2144/01311dd03