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Found 6 matching solutions for this experiment
| Upstream tips |
Protocol tips |
Downstream tips |
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- Incase of Low CYTO-ID Green dye staining in all treatments, increase the reagent concentration (500X dilution of the dye is recommended) and the incubation time.
- Incase of High CYTO-ID Green staining, it could be that the cell culture medium was depleted of nutrients and so change media 4-8 hours before the experiment. |
| Downstream tips |
- Incase of Low CYTO-ID Green dye staining in all treatments, increase the reagent concentration (500X dilution of the dye is recommended) and the incubation time.
- Incase of High CYTO-ID Green staining, it could be that the cell culture medium was depleted of nutrients and so change media 4-8 hours before the experiment. |
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Protocol tips |
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- incubate primary Ab with agitation for 1 hour. |
- If high background or Non-specific bands is present,Try different blocking strategies: Longer blocking times, higher % of blocker, a different blocker, inclusion of blocker protein in antibody dilutions or try a fresh batch of the same blocker. |
| Protocol tips |
| - incubate primary Ab with agitation for 1 hour. |
| Downstream tips |
| - If high background or Non-specific bands is present,Try different blocking strategies: Longer blocking times, higher % of blocker, a different blocker, inclusion of blocker protein in antibody dilutions or try a fresh batch of the same blocker. |
| Upstream tips |
Protocol tips |
Downstream tips |
- Do not freeze component A
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- Incubate the cells overnight (≥16 hours) after adding LC3 reagent.
- For best results, allow cells 48 hours post-transduction for expression levels to equilibrate |
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| Upstream tips |
- Do not freeze component A
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| Protocol tips |
- Incubate the cells overnight (≥16 hours) after adding LC3 reagent.
- For best results, allow cells 48 hours post-transduction for expression levels to equilibrate |
| Upstream tips |
Protocol tips |
Downstream tips |
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- diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween 20 at 4°C with gentle shaking, overnight.
- Dilute Primary Ab at 1:1000 |
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| Protocol tips |
- diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween 20 at 4°C with gentle shaking, overnight.
- Dilute Primary Ab at 1:1000 |
| Upstream tips |
Protocol tips |
Downstream tips |
| - Lyse cells using RIPA mixed with protease inhibitors and phosphatase inhibitors |
- Dilute Primary Ab at 1:500-1:3000 |
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| Upstream tips |
| - Lyse cells using RIPA mixed with protease inhibitors and phosphatase inhibitors |
| Protocol tips |
| - Dilute Primary Ab at 1:500-1:3000 |
| Upstream tips |
Protocol tips |
Downstream tips |
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- 1:500 dilution of primary antibody and overnight incubation at 4°C |
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| Protocol tips |
| - 1:500 dilution of primary antibody and overnight incubation at 4°C |
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