Penicillan Binding Protein Assay青霉素与细胞膜蛋白结合实验
2007-7-4 22:44:00 信息来源:来源网络
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Penicillan Binding Protein Assay青霉素与细胞膜蛋白结合实验
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- Wash cells with 10 mM Tris pH 8
- French press
- Slow speed spin
- High speed spin
- Resuspend in 10 mM Tris
- Sonicate 2 x 15 sec to remove ß-lacatamases
- Wash in 10 mM Tris
- Resuspend in 10 mM Tris
- Store at ?70ºC until needed.
Assay:
- Incubate membranes with ~5 µCi 3H-penicillin at 25ºC for 10 minutes
- Add 1000X cold penicillin
- SDS-PAGE
- Incubate in 1 M sodium salicylate for 30 minutes
- Dry at 80ºC for 2 hours under vacuum
- Expose to photographic film for 3-30 days.
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实验频道录入:ztwpierre 责任编辑:ztwpierre