| Fat Turnover Assay in Drosophila
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Author:
Date:
2016-11-05
[Abstract] Like all animals, Drosophila shows robust fat (triglyceride) turnover, i.e., they synthesize, store and utilize triglyceride for their daily metabolic needs. The protocol describes a simple assay to measure this turnover of triglycerides in Drosophila.
[摘要] 像所有动物一样，果蝇表现出强劲的脂肪（甘油三酯）周转，即，它们合成，储存和利用甘油三酯用于其日常代谢需要。 该方案描述了测量果蝇中甘油三酯转化的简单测定法。 几乎所有的动物都以 糖原和甘油三酯。 许多生理，病理和环境条件引起这些能量储备的总水平的变化，特别是甘油三酯。 然而，并不总是清楚甘油三酯中所导致的变化是由于减少的分解，增加的合成，或反之亦然。 使用此协议，可以确定新合成的甘油三酯在苍蝇中的合成速率和降解。
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| Adoptive Transfer of Memory B Cells
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Author:
Date:
2015-08-20
[Abstract] The adoptive transfer of antigen-specific B cells into mice that cannot recognize that specific antigen has two main advantages. The first is determining exactly when the B cells were transferred and exposed to antigen. The second is that all B cells that can bind that antigen are the ones that were transferred; no new antigen-specific B cells will emerge from the bone marrow. Thus all B cells that were exposed to the antigen and still alive after at least 4 weeks (8 weeks or more is ideal), are memory B cells.
Splenic B cells from B1-8 mice were prepared with an EasySep Mouse B Cell Enrichment Kit according to the manufacturer’s protocol. Single-cell suspensions were transferred intravenously into tail veins of recipient mice. Approximately 1 million NP+ B cells were ...
[摘要] 将抗原特异性B细胞过继转移到不能识别该特异性抗原的小鼠中具有两个主要优点。 第一种是确切地确定B细胞何时转移并暴露于抗原。 第二是可以结合该抗原的所有B细胞是转移的; 从骨髓中不会出现新的抗原特异性B细胞。 因此，暴露于抗原并在至少4周（8周或更长时间是理想的）后仍存活的所有B细胞是记忆B细胞。
根据制造商的方案用EasySep小鼠B细胞富集试剂盒制备来自B1-8小鼠的脾B细胞。 将单细胞悬浮液静脉内转移到受体小鼠的尾静脉中。 每只小鼠转移约100万个NP + B细胞。 转移后约12-24小时，用在明矾中沉淀的50μgNP-CGG腹膜内免疫小鼠。
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