Chromatin Immunoprecipitation: Difference between revisions
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* Alternatively, if multiple immunoprecipitations will be performed from the same chromatin preparation, use the appropriate volume of Protein G Agarose for the correct number of immunoprecipitations. | * Alternatively, if multiple immunoprecipitations will be performed from the same chromatin preparation, use the appropriate volume of Protein G Agarose for the correct number of immunoprecipitations. | ||
20. Incubate for 1 hour at 4°C with rotation. | 20. Incubate for 1 hour at 4°C with rotation. | ||
21. Pellet agarose by brief centrifugation (3000-5000 x g for 1 minute). | 21. Pellet agarose by brief centrifugation (3000-5000 x g for 1 minute). | ||
* Do not spin Protein G Agarose beads at high speeds. Applying excessive g-force may crush or deform the beads and cause them to pellet inconsistently. | * Do not spin Protein G Agarose beads at high speeds. Applying excessive g-force may crush or deform the beads and cause them to pellet inconsistently. | ||
22. Remove 10 μL (1%) of the supernatant as Input and save at 4°C until Section D, step 1. | 22. Remove 10 μL (1%) of the supernatant as Input and save at 4°C until Section D, step 1. | ||