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Found 3 matching solutions for this experiment
| Upstream tips |
Protocol tips |
Downstream tips |
-The 1.25 M glycine must be at room temperature before use.
-The Lysis Buffer must be at room temperature and fully resuspended before use. Vortex briefly to resuspend. |
-For each ChIP reaction, use 10,000–300,000 cells or 0.167–5 mg of tissue.
-1–10 μg of antibody is a typical starting range.
- 10-minute crosslinking step using formaldehyde at a 1% final concentration.
-Keep the cell lysate cooled on ice during sonication. |
|
| Upstream tips |
-The 1.25 M glycine must be at room temperature before use.
-The Lysis Buffer must be at room temperature and fully resuspended before use. Vortex briefly to resuspend. |
| Protocol tips |
-For each ChIP reaction, use 10,000–300,000 cells or 0.167–5 mg of tissue.
-1–10 μg of antibody is a typical starting range.
- 10-minute crosslinking step using formaldehyde at a 1% final concentration.
-Keep the cell lysate cooled on ice during sonication. |
| 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 |
- DO NOT cryo-preserve specimen with sucrose.
-Make fresh 1% formaldehyde before each experiment.
|
-Keep lysate ice-cold. Sonication produces heat, which can denature the chromatin. Allow time between cycles of
sonication to prevent sample overheating.
- DNA fragments should be in 200 -1000 bps in size.
-Add 1-10 µg of immunoprecipitating antibody. |
|
| Upstream tips |
- DO NOT cryo-preserve specimen with sucrose.
-Make fresh 1% formaldehyde before each experiment.
|
| Protocol tips |
-Keep lysate ice-cold. Sonication produces heat, which can denature the chromatin. Allow time between cycles of
sonication to prevent sample overheating.
- DNA fragments should be in 200 -1000 bps in size.
-Add 1-10 µg of immunoprecipitating antibody. |
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