실험 Q&A Immunology > Immunofluorescence
biotin질문요
레벨1 윤지혜
For in-vivo tracking, the platelets were biotinylated using 0.1 mM NHS-biotin and could be later stained with streptavidin conjugated to phycoerytherin (PE) Surface expression of P-selectin was confirmed by flow cytometry using anti-P-selectin-fluorescein isothiocyanate. Biotinylated WT platelets were activated in vitro. The platelets were infused in mice. At the indicated time points, blood was drawn and the infused biotinylated platelets were identified and percentage of P-selectin positive platelets determined by flow cytometry. biotin에 대해 공부하구 있는데 정말 모르겠어요ㅠ 이 논문 내용을 간략히 말씀드리면 활성화된 혈소판표면에 P-selectin이란게 있는데 쥐에 넣어주면 백혈구랑 반응해서 시간이 지나면 P-selectin이 사라지거든요. 쥐한테 P-selectin이 있는 혈소판을 넣어준후 시간이 지나고 넣어준거 중에 P-selectin이 있는 혈소판양을 측정하는건데요. biotin으로 표지해서 제가 넣어준 혈소판만을 알수 있는 거잖아요 biotin에는 PE라는 형광물질이 붙은 streptavidin이 붙고 p-selectin에는 FITC가 붙은 anti-p-selectin이 붙잖아요 그면 PE랑 FITC 혈광 차이로 flow cytometry로 알아보는 건가요??????????
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