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Found 3 matching solutions for this experiment
| Upstream tips |
Protocol tips |
Downstream tips |
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Immunoprecipitations were carried out in 1 ml of IP buffer (20 mM Tris–HCl pH 8.0, 150 mM NaCl, 2 mM EDTA, 1 % Triton-X 100). Three µg antibody was used per 20 µg purified chromatin. Input chromatin was obtained after preclearing by de-crosslinking and purified using the Qiaquick column (Qiagen) according to manufacturer’s instructions. Immunoprecipitations were carried out with inverting at 4 °C for 14–16 h |
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| Protocol tips |
| Immunoprecipitations were carried out in 1 ml of IP buffer (20 mM Tris–HCl pH 8.0, 150 mM NaCl, 2 mM EDTA, 1 % Triton-X 100). Three µg antibody was used per 20 µg purified chromatin. Input chromatin was obtained after preclearing by de-crosslinking and purified using the Qiaquick column (Qiagen) according to manufacturer’s instructions. Immunoprecipitations were carried out with inverting at 4 °C for 14–16 h |
| Upstream tips |
Protocol tips |
Downstream tips |
| -Add protease inhibitors to all lysis solutions before use. |
-Keep cells on ice between the rounds of homogenisations.
- Increase or decrease the homogenization step to maximize the yield of nuclei depending on cell line.
-Do not place ethidium bromide in the agarose gel or the electrophoresis buffer, because of the presence of SDS. |
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| Upstream tips |
| -Add protease inhibitors to all lysis solutions before use. |
| Protocol tips |
-Keep cells on ice between the rounds of homogenisations.
- Increase or decrease the homogenization step to maximize the yield of nuclei depending on cell line.
-Do not place ethidium bromide in the agarose gel or the electrophoresis buffer, because of the presence of SDS. |
| Upstream tips |
Protocol tips |
Downstream tips |
| -It is highly critical that the chromatin is of appropriate size and concentration. |
-It is important to keep the tissue cold to avoid protein degradation.
-Use fresh formaldehyde that is not past the manufacturer's expiration date. |
-Once in solution, store 1M DTT at -20°C. |
| Upstream tips |
| -It is highly critical that the chromatin is of appropriate size and concentration. |
| Protocol tips |
-It is important to keep the tissue cold to avoid protein degradation.
-Use fresh formaldehyde that is not past the manufacturer's expiration date. |
| Downstream tips |
| -Once in solution, store 1M DTT at -20°C. |
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