分析测试百科网

搜索

喜欢作者

微信支付微信支付
×

Penicillan Binding Protein Assay青霉素与细胞膜蛋白结合实验

2019.5.20
头像

zhaochenxu

致力于为分析测试行业奉献终身

  1. Wash cells with 10 mM Tris pH 8

  2. French press

  3. Slow speed spin

  4. High speed spin

  5. Resuspend in 10 mM Tris

  6. Sonicate 2 x 15 sec to remove ß-lacatamases

  7. Wash in 10 mM Tris

  8. Resuspend in 10 mM Tris

  9. Store at ?70ºC until needed.


Assay:

  1. Incubate membranes with ~5 µCi 3H-penicillin at 25ºC for 10 minutes

  2. Add 1000X cold penicillin

  3. SDS-PAGE

  4. Incubate in 1 M sodium salicylate for 30 minutes

  5. Dry at 80ºC for 2 hours under vacuum

  6. Expose to photographic film for 3-30 days.



生物在线
文章推荐