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Array-based ELISAs for high-throughput analysis of human cytokines.
Biotechniques. 2001 Jul; 31(1):186-90, 192-4.B

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.

Authors+Show Affiliations

Pierce Endogen, Inc., 30 Commerce Way, Woburn, MA 01801-1059, USA. mmoody@endogen.comNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

11464511

Citation

Moody, M D., et al. "Array-based ELISAs for High-throughput Analysis of Human Cytokines." BioTechniques, vol. 31, no. 1, 2001, pp. 186-90, 192-4.
Moody MD, Van Arsdell SW, Murphy KP, et al. Array-based ELISAs for high-throughput analysis of human cytokines. BioTechniques. 2001;31(1):186-90, 192-4.
Moody, M. D., Van Arsdell, S. W., Murphy, K. P., Orencole, S. F., & Burns, C. (2001). Array-based ELISAs for high-throughput analysis of human cytokines. BioTechniques, 31(1), 186-90, 192-4.
Moody MD, et al. Array-based ELISAs for High-throughput Analysis of Human Cytokines. BioTechniques. 2001;31(1):186-90, 192-4. PubMed PMID: 11464511.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Array-based ELISAs for high-throughput analysis of human cytokines. AU - Moody,M D, AU - Van Arsdell,S W, AU - Murphy,K P, AU - Orencole,S F, AU - Burns,C, PY - 2001/7/24/pubmed PY - 2002/1/5/medline PY - 2001/7/24/entrez SP - 186-90, 192-4 JF - BioTechniques JO - BioTechniques VL - 31 IS - 1 N2 - 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. SN - 0736-6205 UR - https://www.unboundmedicine.com/medline/citation/11464511/Array_based_ELISAs_for_high_throughput_analysis_of_human_cytokines_ L2 - https://www.future-science.com/doi/full/10.2144/01311dd03?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub=pubmed DB - PRIME DP - Unbound Medicine ER -