Purification of murine macrophage cytotoxin (MCT).
Macrophage cytotoxin (MCT) can be induced from peritoneal exudate macrophage monolayers (PEMM) obtained from thioglycollate-treated mice, by exposure of PEMM to lipopolysaccharide (LPS) or to double-stranded polyinosinic: poly-cytidylic acid (poly-l:poly-C). MCT is highly labile even upon storage at 4 degrees C, and is irreversibly denatured by isolectricfocusing, polyacrylamide gel electrophoresis in the absence of sodium dodecyl sulfate (SDS), or by exposure to ethylene glycol. alpha-MCT [150,000 daltons (d)] has been highly purified (2,000- to 5,000-fold) from serum-free, PEMM supernatants by a scheme of concentration, molecular sieving on Ultrogel AcA 44, negative hydrophobic affinity chromatography on benzyl-agarose, and ion exchange chromatography on aminoethyl-agarose. The scheme results in high yield of MCT, in part because of the rapidity with which the labile toxin is manipulated due to the tandemization of the chromatographic steps.
D000818:Animals; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003603:Cytotoxins / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D008264:Macrophages; D051379:Mice; D008810:Mice, Inbred C57BL; D013696:Temperature
D003603:Cytotoxins