Abstract
ELISAs offer excellent specificity and, once fully optimized, sensitivity that rivals that of bioassays. The major variables that need to be experimentally determined when developing an ELISA are the optimal number of fresh cells required per well, the optimal antigen concentrations for stimulation, period of culture, and the anticipated intensity of the response. In this chapter, we review the major factors to be considered in the development and application of ultrasensitive ELISAs to the analysis of human immune responses. We specify the conditions we have found to be optimal for quantifying a number of cytokines of demonstrated relevance to human immune regulation and discuss the major pitfalls inherent in this approach.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2008 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Stefura, W.P. et al. (2008). Ultrasensitive ELISA for Measurement of Human Cytokine Responses in Primary Culture. In: Jones, M.G., Lympany, P. (eds) Allergy Methods and Protocols. Methods in Molecular Medicine, vol 138. Humana Press. https://doi.org/10.1007/978-1-59745-366-0_10
Download citation
DOI: https://doi.org/10.1007/978-1-59745-366-0_10
Publisher Name: Humana Press
Print ISBN: 978-0-89603-896-7
Online ISBN: 978-1-59745-366-0
eBook Packages: Springer Protocols