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文献和实验Intracellular Cytokine Staining Protocol
stimulation period for induction of a given cytokine is variable and has to be determined. For example, the best time for detection of IL-6-producing cells by human LPS-activated monocytes is 6 hours, whereas IL-10 needs at least 24 hours stimulation
Intracellular Cytokine Staining Protocol
. Note: Optimal stimulation period for induction of a given cytokine is variable and has to be determined. For example, the best time for detection of IL-6-producing cells by human LPS-activated monocytes is 6 hours, whereas IL-10 needs at least 24
1、分别加入1/10体积3mol/l NaAc(pH4.5)和等体积5:1酸性酚氯仿溶液,冰浴20min 4℃,11000*g,20min离心。2、吸取上清至一50ml离心管,加入等体积异丙醇,混匀后-20℃放1小时。4、 4℃,1100*g,20min离心,弃上清。5、每管加入3毫升的溶液D,溶解沉淀,再用等体积的酚氯仿各抽提一次。6、加等体积的异丙醇,-20℃沉淀2小时。4℃,10000*g离心20min,弃上清。7、加入10ml冰预冷的75%乙醇,洗涤沉淀2次,晾干沉淀。8、溶解和比色
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