A method for measuring multiple cytokines from small samples.
Commercially available enzyme-linked immunosorbent assay (ELISA) kits are commonly used to assess levels of proinflammatory cytokines in biological samples. Most of these kits require sample volumes of at least 50 microl. Thus, in order to examine multiple cytokines, volumes greater than 100 microl must be collected. However, the volume of many biological samples, especially those collected from the central nervous system (i.e., microdialysates, push-pull perfusions, or cerebrospinal fluid samples), is much less than 100 microl. Therefore, we developed a method for analyzing multiple cytokines from a single, low-volume biological sample, which involves serially assaying the samples on multiple proinflammatory cytokine ELISA kits. In many cases, assaying for one cytokine does not interfere with subsequent assay for another cytokine in the same sample. Moreover, when interference is observed, the interfering factor can be identified and its effect minimized.
O'Connor
Kevin A
KA
Department of Psychology and Center for Neuroscience, University of Colorado, Boulder, CO 80309, USA.
Holguin
Adelina
A
Hansen
Michael K
MK
Maier
Steven F
SF
Watkins
Linda R
LR
D000818:Animals; D001826:Body Fluids / Q000737:chemistry*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D007375:Interleukin-1 / Q000032:analysis*; D015850:Interleukin-6 / Q000032:analysis*; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D018401:Sample Size; D012680:Sensitivity and Specificity; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis*
D007375:Interleukin-1; D015850:Interleukin-6; D014409:Tumor Necrosis Factor-alpha