Method for the isolation of highly purified Salmonella flagellins.
Ten different Salmonella serotypes were grown in a chemically defined medium supplemented with 0.01% yeast extract. After sedimentation of the cells by centrifugation, flagella were detached by exposure to pH 2 for 30 min at room temperature. The flagellaless cells were removed by centrifugation, and the flagellin in the supernatant was further purified by high-speed centrifugation, ammonium sulfate precipitation, and dialysis in 50,000-molecular-weight-cutoff tubing. The 10 flagellin preparations were of a high degree of purity, as demonstrated by electron microscopy, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and measurement of salmonella H and O agglutination titers of antisera raised in rabbits with the flagellin preparations as immunogens.
D000942:Antigens, Bacterial / Q000302:isolation & purification; D001426:Bacterial Proteins / Q000302:isolation & purification*; D005407:Flagella / Q000032:analysis*; D005408:Flagellin / Q000276:immunology / Q000302:isolation & purification*; D008854:Microscopy, Electron; D008970:Molecular Weight; D012475:Salmonella / Q000032:analysis* / Q000145:classification / Q000276:immunology; D012703:Serotyping
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D005408:Flagellin