Procedures were refined and improved for the purification of cytochrome c from Neurospora crassa, for development of specific antisera to the protein, and for immunochemical identification of cytochrome c after in vivo synthesis and radiolabeling with 3H and 35Fe. The cytochrome c was obtained in high yields via column chromatography and salt fractionation, and the product was pure as judged by spectrophotometric and electrophoretic criteria. Effective, specific antisera to cytochrome c could be obtained only after the monomeric protein antigen was crosslinked with glutaraldehyde; immunoprecipitation of cytochrome c synthesized in vivo was achieved efficiently with a double-antibody procedure.
Bibliographical noteFunding Information:
This research was supported by NIH Research Grant GM-19398 from the National Institute of General Medical Sciences.
- Neurospora crassa
- cytochrome c, immunochemical identification, purification
- double-antibody procedure