Protocol tips
Upstream tips |
Protocol tips |
Downstream tips |
-Do not re-freeze the Protein G magnetic beads, the beads should be stored at 4C. |
-Always use ChIP validated antibody.
-Optimal use of 1-3 ug of antibody. |
-The diluted Proteinase K stop solution can not be stored. |
Upstream tips |
-Do not re-freeze the Protein G magnetic beads, the beads should be stored at 4C. |
Protocol tips |
-Always use ChIP validated antibody.
-Optimal use of 1-3 ug of antibody. |
Downstream tips |
-The diluted Proteinase K stop solution can not be stored. |
Publication protocol
Chromatin immunoprecipitation (ChIP) was carried out as previously described using ChIP-IT Express Kit from Active Motif (53008; Carlsbad, CA) [50]. For each ChIP reaction, 20 μg of total chromatin was incubated with 20 μl of Protein G magnetic beads and 1 μg of FLAG-M2 antibody (F3165; Sigma) at 4°C overnight. Both input and precipitated DNA were purified with QIAamp DNA Mini Kit (Qiagen, Valencia, CA) for subsequent quantitative real-time PCR (ChIP-qPCR). The relative enrichment of precipitated DNA was normalized to normal mouse IgG.
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Manufacturer protocol
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