No discussions found
Start your discussion
Share your thoughts or question with experts in your field
Start a discussion
Found 3 matching solutions for this experiment
| Upstream tips |
Protocol tips |
Downstream tips |
| -Do not use formaldehyde if white precipitate is visible in the solution. |
-Use ChIP validated antibody.
- 1X104 cells per ChIP when using high affinity antibodies directed towards more abundant proteins.
-Protease Inhibitor Cocktail III contains DMSO and will remain frozen below 18.4°C.
-Keep cell lysate ice cold. Sonication produces heat, which can denature the chromatin. Allow at least 30
seconds between cycles of sonication to prevent sample overheating.
|
|
| Upstream tips |
| -Do not use formaldehyde if white precipitate is visible in the solution. |
| Protocol tips |
-Use ChIP validated antibody.
- 1X104 cells per ChIP when using high affinity antibodies directed towards more abundant proteins.
-Protease Inhibitor Cocktail III contains DMSO and will remain frozen below 18.4°C.
-Keep cell lysate ice cold. Sonication produces heat, which can denature the chromatin. Allow at least 30
seconds between cycles of sonication to prevent sample overheating.
|
| Upstream tips |
Protocol tips |
Downstream tips |
|
-Just because an antibody works well in a Western blot does not always indicate it will perform well in chromatin immunoprecipitation.
Unlike a Western blot that detects proteins that have been denatured, a ChIP antibody must recognize the target protein in its native state. Use ChIP-validated antibody.
-Use 2-10 μg of your ChIP antibody depending on the abundance of your protein target. |
|
| Protocol tips |
-Just because an antibody works well in a Western blot does not always indicate it will perform well in chromatin immunoprecipitation.
Unlike a Western blot that detects proteins that have been denatured, a ChIP antibody must recognize the target protein in its native state. Use ChIP-validated antibody.
-Use 2-10 μg of your ChIP antibody depending on the abundance of your protein target. |
| Upstream tips |
Protocol tips |
Downstream tips |
| -Once in solution, store 1M DTT at -20°C. |
-For optimal ChIP results, use approximately 4 X 106 cells for each immunoprecipitation to be performed.
-For optimal ChIP results, it is highly critical that the chromatin is of appropriate size and concentration.
-For optimal ChIP results, use approximately 5 to 10 µg of digested, cross-linked chromatin per immunoprecipitation. |
|
| Upstream tips |
| -Once in solution, store 1M DTT at -20°C. |
| Protocol tips |
-For optimal ChIP results, use approximately 4 X 106 cells for each immunoprecipitation to be performed.
-For optimal ChIP results, it is highly critical that the chromatin is of appropriate size and concentration.
-For optimal ChIP results, use approximately 5 to 10 µg of digested, cross-linked chromatin per immunoprecipitation. |
Can't find the product you've used to perform this experiment? It would be great if you can help us by
Adding a product!