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Found 15 matching solutions for this experiment
| Upstream tips |
Protocol tips |
Downstream tips |
| This kit uses a green fluorescent probe that reacts with autophagic vacuoles and is detectable by flow cytometry |
Resuspend 5 × 10*5 cells in 1× assay buffer and incubate with the Cyto‐ID dye at 37 ºC for 30 minutes in the dark, followed by flow‐cytometric analysis. For Fluorescence microscopy, grow the cells on coverslips after respective treatment incubate them with the dual detection reagent (Cyto‐ID dye and Hoechst 3342 nuclear stain) at 37 ºC for 30 minutes in the dark. |
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. |
| Upstream tips |
| This kit uses a green fluorescent probe that reacts with autophagic vacuoles and is detectable by flow cytometry |
| Protocol tips |
| Resuspend 5 × 10*5 cells in 1× assay buffer and incubate with the Cyto‐ID dye at 37 ºC for 30 minutes in the dark, followed by flow‐cytometric analysis. For Fluorescence microscopy, grow the cells on coverslips after respective treatment incubate them with the dual detection reagent (Cyto‐ID dye and Hoechst 3342 nuclear stain) at 37 ºC for 30 minutes in the dark. |
| 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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Downstream tips |
| Lyse cells in ice-cold mammalian lysis buffer |
For WB, incubate with primary antibody diluted to 2 ug/ml in total volume of 3 mL in Blocking
buffer for one hour at room temperature.
For IF, stain with primary antibody for 45 min at RT in 40 ml of 1%BSA-PBS by forming a
drop on the coverslip |
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| Upstream tips |
| Lyse cells in ice-cold mammalian lysis buffer |
| Protocol tips |
For WB, incubate with primary antibody diluted to 2 ug/ml in total volume of 3 mL in Blocking
buffer for one hour at room temperature.
For IF, stain with primary antibody for 45 min at RT in 40 ml of 1%BSA-PBS by forming a
drop on the coverslip |
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For WB, dilute primary Ab at 1:1000 and for immunoprecipitation at 1:100.
For western blots, incubate membrane with diluted antibody in 5% w/v BSA, 1X TBS,
0.1% Tween®20 at 4°C with gentle shaking, overnight |
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| Protocol tips |
For WB, dilute primary Ab at 1:1000 and for immunoprecipitation at 1:100.
For western blots, incubate membrane with diluted antibody in 5% w/v BSA, 1X TBS,
0.1% Tween®20 at 4°C with gentle shaking, overnight |
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For Western Blotting (starting dilution 1:100, dilution range 1:100-1:1000.
For Immunofluorescent staining incubate with primary antibody at 8°C overnight. |
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| Protocol tips |
For Western Blotting (starting dilution 1:100, dilution range 1:100-1:1000.
For Immunofluorescent staining incubate with primary antibody at 8°C overnight. |
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Dilute primary antibody at 1:1,000 dilution and incubate overnight at 4°C |
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| Protocol tips |
| Dilute primary antibody at 1:1,000 dilution and incubate overnight at 4°C |
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Dilute primary Ab between 1:500 - 1:1000 and incubate overnight at 4°C |
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| Protocol tips |
| Dilute primary Ab between 1:500 - 1:1000 and incubate overnight at 4°C |
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Dilute primary Ab between 1:500 - 1:1000 and incubate overnight at 4°C |
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| Protocol tips |
| Dilute primary Ab between 1:500 - 1:1000 and incubate overnight at 4°C |
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Dilute primary Ab at 1:2000 and incubate at overnight at 4°C. |
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| Protocol tips |
| Dilute primary Ab at 1:2000 and incubate at overnight at 4°C. |
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Dilute primary Ab at 1:1000 and incubate overnight at 4°C |
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| Protocol tips |
| Dilute primary Ab at 1:1000 and incubate overnight at 4°C |
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Dilute primary Ab at 1:1000 and incubate overnight at 4°C |
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| Protocol tips |
| Dilute primary Ab at 1:1000 and incubate overnight at 4°C |
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Downstream tips |
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Dilute primary Ab at 1:1000 and incubate overnight at 4°C |
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| Protocol tips |
| Dilute primary Ab at 1:1000 and incubate overnight at 4°C |
| Upstream tips |
Protocol tips |
Downstream tips |
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Dilute primary Ab at 1:1000 and incubate overnight at 4°C |
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| Protocol tips |
| Dilute primary Ab at 1:1000 and incubate overnight at 4°C |
| Upstream tips |
Protocol tips |
Downstream tips |
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Dilute primary Ab at 1:1000 and incubate overnight at 4°C |
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| Protocol tips |
| Dilute primary Ab at 1:1000 and incubate overnight at 4°C |
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For WB, dilute primary Ab between 1:10000 - 1:50000 |
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| Protocol tips |
| For WB, dilute primary Ab between 1:10000 - 1:50000 |
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Use a concentration of 1 - 5 µg/ml. |
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| Protocol tips |
| Use a concentration of 1 - 5 µg/ml. |
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