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Found 3 matching solutions for this experiment
| Upstream tips |
Protocol tips |
Downstream tips |
-If any reagent has formed a precipitate, warm at 55 °C until dissolved. Allow to equilibrate to room temperature before use.
|
-Use 1 µg of antibody per well.
-The range of cells that can be used with this kit is 0.2–2.5 x 105/well/ChIP sample. Using more than 0.5 million cells per well will increase the non-specific binding and reduce the yield and specificity of the ChIP reaction. If higher yield of ChIP DNA is desired for downstream applications (e.g. ChIP-Seq., ChIP-chip): a) DNA could be pooled by eluting consecutively (with 50 µl of elution buffer) from multiple individual ChIP reactions. |
|
| Upstream tips |
-If any reagent has formed a precipitate, warm at 55 °C until dissolved. Allow to equilibrate to room temperature before use.
|
| Protocol tips |
-Use 1 µg of antibody per well.
-The range of cells that can be used with this kit is 0.2–2.5 x 105/well/ChIP sample. Using more than 0.5 million cells per well will increase the non-specific binding and reduce the yield and specificity of the ChIP reaction. If higher yield of ChIP DNA is desired for downstream applications (e.g. ChIP-Seq., ChIP-chip): a) DNA could be pooled by eluting consecutively (with 50 µl of elution buffer) from multiple individual ChIP reactions. |
| Upstream tips |
Protocol tips |
Downstream tips |
|
DNA-protein cross-linking was carried out by addition of 11.1% formaldehyde and incubation for 5 min @ 25°C. Cells were lysed by addition of Lysis Buffer A. The lysates were sonicated @ 6°C; and the concentration of sheared DNA was measured by NanoDrop. Four µg of ERα antibody (AER304, mouse monoclonal, Millipore) or mouse non-immune IgG (Millipore) was utilized for IPs. |
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| Protocol tips |
| DNA-protein cross-linking was carried out by addition of 11.1% formaldehyde and incubation for 5 min @ 25°C. Cells were lysed by addition of Lysis Buffer A. The lysates were sonicated @ 6°C; and the concentration of sheared DNA was measured by NanoDrop. Four µg of ERα antibody (AER304, mouse monoclonal, Millipore) or mouse non-immune IgG (Millipore) was utilized for IPs. |
| 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. |
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