Abstract
Here we describe the methods for the expression of human and murine soluble tumor necrosis factor (hTNF and mTNF) in Escherichia coli cells and for their extraction, purification, and characterization. The expression and purification procedure takes about 2 wk. Human and murine TNF can be purified from crude extracts with high yields (>50 mg from 1 L of fermentation) by hydrophobic-interaction chromatography, ion-exchange chromatography, and gel-filtration chromatography. The purity and the identity of the final products can be checked by SDS-PAGE, ELISA, Western blot, analytical gel-filtration chromatography, mass spectrometry, and lipopolysaccharide assay. The biological activity of both human and murine TNF is assessed by in vitro cytolytic assays using murine L-M cells. Purified hTNF and mTNF can be used for in vitro and in vivo studies in animal models.
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
