Application of Globulol
A technology of cytokines and compositions, applied in the application field of Globuol, can solve the problems of affecting, reducing the quality of life of patients, and threatening life.
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Embodiment 1
[0029] Embodiment 1MTT experiment
[0030] MTT assay was used to determine the combined experiment of cytokines γ-IFN, TNF-α and IL-6 and the influence of Globulol on the growth of BEAS-2B cells. The specific implementation plan is as follows:
[0031] 1. Take the BEAS-2B cells in a good growth state, count them with a cell counting board after blowing down, and adjust the final concentration of cells to 5×10 4 pieces / ml. Add 100 μl of cell suspension per well in a 96-well plate at 37 °C and 5% CO 2 After culturing in the incubator for 24 hours, discard the old medium;
[0032] 2. Set up three groups of experiments, respectively (1) set the concentration of γ-IFN, TNF-α and IL-6 to 100ng / ml, 60ng / ml and 10ng / ml respectively, (2) set the concentration of Globulol to 2μM, 5μM, 10μM, 20μM, (3) set cytokine combination (γ-IFN 100ng / ml, TNF-α60ng / ml, IL-6 10ng / ml) in combination with different concentrations of Globulol (2μM, 5μM, 10μM, 20μM). Set up 4 replicate wells for each ...
Embodiment 2
[0034] Embodiment 2 trypan blue experiment
[0035] Use the trypan blue experiment to measure the influence of the combined use of cytokines γ-IFN, TNF-α and IL-6 on the proliferation of BEAS-2B cells. The specific implementation is as follows:
[0036] 1. Take the BEAS-2B cells in good growth state, count them with a cell counting board after blowing down, and adjust the final concentration of cells to 0.2×10 5 pieces / ml. Add 2ml of cell suspension per well in a 35mm cell culture dish and place at 37°C and 5% CO 2 After culturing in the incubator for 24 h, the old medium was discarded.
[0037] 2. Set up two groups of experiments respectively, respectively (1) set the concentrations of γ-IFN, TNF-α and IL-6 as 100ng / ml, 60ng / ml and 10ng / ml respectively, (2) set the combination of cytokines (γ- IFN 100ng / ml, TNF-α 60ng / ml, IL-6 10ng / ml) were used in combination with different concentrations of Globulol (2μM, 5μM, 10μM, 20μM), and 4 replicate wells were set for each concentr...
Embodiment 3
[0039] Embodiment 3 Caspase-1 activity experiment
[0040] Take the BEAS-2B cells in good growth state, count them with a cell counting board after blowing down, and adjust the final concentration of cells to 0.2×10 5 pieces / ml. Add 5ml of cell suspension to a 60mm cell culture dish, place it in an incubator at 37°C and 5% CO2 for 24 hours, and then discard the old medium.
[0041] Set up two groups of experiments respectively, respectively set (1) γ-IFN, TNF-α and IL-6 concentrations are 100ng / ml, 60ng / ml and 10ng / ml respectively (2) set up cytokine combination (γ-IFN 100ng / ml , TNF-α60ng / ml, IL-610ng / ml) combined with different concentrations of Globulol (2μM, 5μM, 10μM, 20μM). After cell culture for 72 hours, trypsinize the adherent cells, centrifuge at 600g at 4°C for 5 minutes to collect the cells, carefully aspirate the supernatant while ensuring that no cells are aspirated as much as possible, and wash once with PBS. After aspirating the supernatant as before, add th...
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