Ultrasensitive ELISA for Measurement of Human Cytokine Responses in Primary Culture
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.
- 缺陷噬菌体(defective phage)
- Titration of retrovirus
- Two-Dimensional Polyacrylamide Gel Electrophoresis
- Qualitative and Quantitative Determination of Quorum Sensing Inhibition In Vitro
- Bovine Leukemia Virus Ribozymes
- Detection of Clostridium botulinum by Multiplex PCR in Foods and Feces
- Isolation of Human Tonsillar Dendritic Cells
- Isolation of Human Skin Dendritic Cells by In Vitro Migration
- Measurement of Cytokines in Cerebrospinal Fluid
- Titration of Reagents