Protocol tips
| Upstream tips |
Protocol tips |
Downstream tips |
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-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. |
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| 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. |
Publication protocol
An EZ‐ChIP™ Assay Kit (Cat#17–371; Millipore) was used to perform quantitative chromatin immunoprecipitation (qChIP) assays according to the manufacturer's protocol. In brief, cells were treated with 37% formaldehyde to crosslink proteins, and then the reaction was terminated with 0.125 M glycine. After sonication, chromatin–protein complexes were immunoprecipitated with 5 μg of anti‐Sall4 antibody (sc‐101147; Santa Cruz). Regions of interest were amplified in triplicate from precipitated samples by real‐time PCR, and the amount of precipitated DNA was calculated as a percentage of the input sample.
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Papers
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Manufacturer protocol
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