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文献和实验ChIP protocol for X. laevis Lens1/FoxE3 enhancer
) and incubate for 30 min at 37°C. (41) Add 4 µl 0.5M EDTA, 8 µl 1M Tris-HCl, and 1 µl Proteinase K (EZ ChIP kit) and incubate at 45°C for 2 hours. (42) Purify DNA using Spin columns in EZ ChIP kit or Qiagen PCR purification kit. (43) The purified
Preparation of Brain Membrane Fractions
in approximately 5 ml water. Add sucrose and water to give a final volume of 45 ml 1.2 M sucrose, add HEPES to 5 mM and the protease inhibitors as above. Homogenize in glass FEP polymer-homogenizer (3 strokes, 1000 rpm).Spin at 9200 x g for 20 min.Resuspend pellets
[分享] Strip Antibodies and Re-use Protein blots membrane
10min in PBST 5. Transfer the membrane to strip buffer (preheated to 55-65C) for 20min, 6. Repeat step 5 once 7. Rinse 3X in PBST followed by 10min in PBST 8. Check strip efficiency (normally, it is OK, but if the expression is too high
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