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发表于 2016-10-11 20:35:05
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Application Notes
Recommended Assay Procedure:
Procedure for Using Leukocyte Activation Cocktail, with BD GolgiPlugTM : Rapidly thaw the cocktail at 37°C in a water bath and add 2 μL of cocktail for every 1 mL of cell culture (e.g., ~10^6 cells/mL) and mix thoroughly. Place culture in a 37°C humidified CO2 incubator for 4-6 hr. Following activation, harvest and wash cells with FACS Staining Buffer (e.g., BD PharmingenTM Stain Buffer, with FBS, Cat. No. 554656) for use in antibody staining protocols. Treatment of stimulated cells for 4 to 6 hours with Leukocyte Activation Cocktail, with BD GolgiPlug significantly increases the ability to detect cytokine-producing cells by immunofluorescent staining. It is recommended that Leukocyte Activation Cocktail, with BD GolgiPlug not be kept in cell culture for longer than 12 hours. BD GolgiPlug been found to have differential effects on intracellular cytokine staining that is time, activator and cytokine dependent, therefore the need to activate cells for periods longer than 4-6 hr may be determined empirically by the investigator. |
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发表于 2016-9-23 08:22:34
发表于 2016-10-11 20:20:33
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