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Found 3 matching solutions for this experiment
| Upstream tips |
Protocol tips |
Downstream tips |
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hPDLCs were rinsed with PBS for 3 times and fixed in 4% paraformaldehyde for 20 min. The fixed cells were treated with 0.5% Triton X-100 and 3% hydrogen peroxide for 15 min. The cells were then incubated for 1 h at 37 °C with a primary mouse anti-pan-cytokeratin antibody (1:500) |
Thereafter, the cells were washed with PBS for 3 times and incubated with corresponding secondary antibodies for 30 min at 37 °C. Finally, the cells were stained with a DAB kit (ZSGB-BIO, Beijing, China) and haematoxylin and viewed under an inverted microscope (Olympus, IX71, Tokyo, Japan). |
| Protocol tips |
| hPDLCs were rinsed with PBS for 3 times and fixed in 4% paraformaldehyde for 20 min. The fixed cells were treated with 0.5% Triton X-100 and 3% hydrogen peroxide for 15 min. The cells were then incubated for 1 h at 37 °C with a primary mouse anti-pan-cytokeratin antibody (1:500) |
| Downstream tips |
| Thereafter, the cells were washed with PBS for 3 times and incubated with corresponding secondary antibodies for 30 min at 37 °C. Finally, the cells were stained with a DAB kit (ZSGB-BIO, Beijing, China) and haematoxylin and viewed under an inverted microscope (Olympus, IX71, Tokyo, Japan). |
| Upstream tips |
Protocol tips |
Downstream tips |
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. One million cells were incubated with 100 μl of MAb mixture directed against keratin overnight at 4°C. Cells were washed and incubated with 100 μl of premixed subclass-specific secondary antibody. DNA was labelled with PI and RNase (Sigma-Aldrich) treated (analyzed using a FACSCalibur flow cytometer, BD Biosciences, San Jose, CA) or DAPI (analyzed using an LSRII flow cytometer BD Biosciences). |
A minimum of 30,000 single cell events were collected for each sample. |
| Protocol tips |
| . One million cells were incubated with 100 μl of MAb mixture directed against keratin overnight at 4°C. Cells were washed and incubated with 100 μl of premixed subclass-specific secondary antibody. DNA was labelled with PI and RNase (Sigma-Aldrich) treated (analyzed using a FACSCalibur flow cytometer, BD Biosciences, San Jose, CA) or DAPI (analyzed using an LSRII flow cytometer BD Biosciences). |
| Downstream tips |
| A minimum of 30,000 single cell events were collected for each sample. |
| Upstream tips |
Protocol tips |
Downstream tips |
|
One million cells were incubated with 100 μl of MAb mixture directed against keratin overnight at 4°C. Cells were washed and incubated with 100 μl of premixed subclass-specific secondary antibody. DNA was labelled with PI and RNase (Sigma-Aldrich) treated (analyzed using a FACSCalibur flow cytometer, BD Biosciences, San Jose, CA) or DAPI (analyzed using an LSRII flow cytometer BD Biosciences). |
A minimum of 30,000 single cell events were collected for each sample. |
| Protocol tips |
| One million cells were incubated with 100 μl of MAb mixture directed against keratin overnight at 4°C. Cells were washed and incubated with 100 μl of premixed subclass-specific secondary antibody. DNA was labelled with PI and RNase (Sigma-Aldrich) treated (analyzed using a FACSCalibur flow cytometer, BD Biosciences, San Jose, CA) or DAPI (analyzed using an LSRII flow cytometer BD Biosciences). |
| Downstream tips |
| A minimum of 30,000 single cell events were collected for each sample. |
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