No discussions found
Start your discussion
Share your thoughts or question with experts in your field
Start a discussion
Found 3 matching solutions for this experiment
Upstream tips |
Protocol tips |
Downstream tips |
|
For measuring intracellular cytokines, PBMCs were pre-treated for 15 min with the hydroxamic acid derivative GM6001 (100 μM, CALBIOCHEM) to inhibit the shedding of IL-6Rα (4) and stimulated for 4 hours with or without phorbol myristate acetate (50ng/ml, PMA) and ionomycin (1 μg/ml) (both from Sigma Aldrich) in the presence of Golgiplug (BD Bioscience |
In most cases, greater than 100,000 events were collected for analysis. Collected data were analyzed using FlowJo software (Tree Star). |
Protocol tips |
For measuring intracellular cytokines, PBMCs were pre-treated for 15 min with the hydroxamic acid derivative GM6001 (100 μM, CALBIOCHEM) to inhibit the shedding of IL-6Rα (4) and stimulated for 4 hours with or without phorbol myristate acetate (50ng/ml, PMA) and ionomycin (1 μg/ml) (both from Sigma Aldrich) in the presence of Golgiplug (BD Bioscience |
Downstream tips |
In most cases, greater than 100,000 events were collected for analysis. Collected data were analyzed using FlowJo software (Tree Star). |
Upstream tips |
Protocol tips |
Downstream tips |
|
>20 × 105 disaggregated sphere cells were stained with a stem cell‐specific antibody panel and analysed on a BDTM LSR II flow cytometer. To a volume of 100 μL cell suspension, 5 μL antibody mix was added, mixed and incubated for 20 minutes at 4°C in the dark and washed twice with 250 μL DPBS or FACS buffer. |
|
Protocol tips |
>20 × 105 disaggregated sphere cells were stained with a stem cell‐specific antibody panel and analysed on a BDTM LSR II flow cytometer. To a volume of 100 μL cell suspension, 5 μL antibody mix was added, mixed and incubated for 20 minutes at 4°C in the dark and washed twice with 250 μL DPBS or FACS buffer. |
Upstream tips |
Protocol tips |
Downstream tips |
|
Mononuclear cells (MNCs) were separated by Ficoll-Hypaque density gradient centrifugation. Immunostaining was performed by a standard indirect immunofluorescence method, as previously described |
|
Protocol tips |
Mononuclear cells (MNCs) were separated by Ficoll-Hypaque density gradient centrifugation. Immunostaining was performed by a standard indirect immunofluorescence method, as previously described |
Can't find the product you've used to perform this experiment? It would be great if you can help us by
Adding a product!