Tumour-necrosis factor from the rabbit. IV. Purification and chemical characterization.
Serum from rabbits with BCG/endotoxin-induced shock is growth inhibitory or cytotoxic to a range ot tumour cell lines. The active component, tumour-necrosis factor (TNF), has been purified 1000-fold by sequential salt precipitation, ion-exchange chromatography and gel-filtration. TNF had a mol. wt of 67,000 on gradient PAGE and 39,000 on Ultrogel AcA44 gel-filtration. The isoelectric point was pH 5.1-5.2. TNF was susceptible to the proteolytic enzyme pronase, but resistant to trypsin or papain. On isopycnic ultracentrifugation it had a buoyant density of 1.27, confirming that it is protein in nature, with little or no carbohydrate. This is also suggested by its failure to bind to a range of lectins.
D000818:Animals; D002460:Cell Line; D002500:Centrifugation, Isopycnic; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000302:isolation & purification*; D007526:Isoelectric Point; D008970:Molecular Weight; D009336:Necrosis / Q000473:pathology*; D009374:Neoplasms, Experimental / Q000473:pathology*; D011817:Rabbits; D014409:Tumor Necrosis Factor-alpha
D006023:Glycoproteins; D014409:Tumor Necrosis Factor-alpha