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| Cell Lysates For Western Blotting
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| [ 文章来源： | 文章作者：
| 发布时间：2006-09-27|
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Cell Lysates For Western Blotting
Reagents / Solutions Lysis Buffer:
10ml 10% Sodium dodecyl sulphate (SDS)
10ml Glycerol
10ml b-mercaptoethanol
8ml 0.5M Tris pH6.8
1ml 0.1% bromophenol blue
51ml H2O
Protocol
- Spin down 106 cells and wash, if required, in phosphate buffered saline.
- Resuspend cells in 100µl warmed lysis buffer, vortex then boil for 5 - 10 minutes to break up DNA.
- Cool (not on ice - the SDS will precipitate) and centrifuge to collect droplets.
- Vortex briefly to mix.
- The samples may now be analysed by SDS - PAGE and Western Blotting, or
- Store samples at -20°C until required.
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Western Blots