PMID: 2125621Dec 31, 1990Paper

Effect of anticoagulants and heat on the detection of tumor necrosis factor in murine blood

Journal of Immunological Methods
P A Holobaugh, D C McChesney

Abstract

Assays for tumor necrosis factor (TNF) may be inhibited by nonspecific factors present in body fluids. We found that the ability to quantitate TNF is greatly affected by blood processing methods. Mice were anesthetized by inhalation of methoxyflurane before obtaining blood by cardiac puncture. The blood from a group of mice was allowed to clot before recovery of serum. Plasmas were obtained from three other groups of mice after collection of blood into tubes containing EDTA, sodium citrate, or preservative-free heparin. The pooled serum or plasmas were spiked with rhTNF to a final concentration of 1 microgram/ml and aliquoted for frozen storage. The serum and plasmas were divided into heated (56 degrees C for 30 min) and unheated portions prior to a standard L929 cytotoxicity assay. Comparison of absorbances at 595 nm after crystal violet staining of cells revealed differences in detection of TNF in plasmas compared to serum and in heated compared to nonheated samples. Citrated plasma clotted in the assay at dilutions at or below 1:25. EDTA plasma consistently produced unexplained lysis of L929 cells in both heated and unheated unspiked samples. We conclude that TNF levels should be determined only in heat treated serum samples...Continue Reading

References

Jan 6, 1989·Journal of Immunological Methods·A MeagerJ Woolley
Nov 6, 1986·Journal of Immunological Methods·S M Kramer, M E Carver

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Citations

Nov 1, 1993·Journal of Bone and Mineral Research : the Official Journal of the American Society for Bone and Mineral Research·J A RianchoJ González-Macías
Oct 20, 1999·Methods : a Companion to Methods in Enzymology·R V House
Jan 1, 2012·Molecular Metabolism·Maximilian BielohubyMartin Bidlingmaier
Jan 24, 2007·Journal of Immunoassay & Immunochemistry·Aysegul YucelAybay Cemalettin
Aug 13, 1999·The American Journal of Physiology·M R BergmanB J Holycross
Jan 14, 1999·The American Journal of Physiology·C A RichardD J Loegering

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