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Author:
Date:
2013-06-20
[Abstract] The LC3B protein plays a critical role in autophagy. Normally, this protein resides in the cytosol, but following cleavage and lipidation with phosphatidylethanolamine, LC3B associates with the phagophore. This localization can be used as a general marker for autophagic membranes. To visualize the LC3B, we used the LC3B Antibody Kit for Autophagy (Invitrogen). As this kit was designed to work with cell coming from cell culture it was not possible to perform it on compact tissues such as nerve cord or ovaries. We thus adapted LC3B labeling initial protocol to be able to label autophagic membranes of adipocytes from terrestrial isopods which form a loose tissue that can be more easily penetrated by antibodies. The following protocol permits a visualization of the autophagic vesicles ...
[摘要] LC3B蛋白在自噬中起关键作用。 通常，这种蛋白质存在于细胞质中，但是在用磷脂酰乙醇胺切割和脂化后，LC3B与吞噬细胞结合。 这种定位可以用作自噬膜的一般标记。 为了显现LC3B，我们使用用于自噬的LC3B抗体试剂盒（Invitrogen）。 因为该试剂盒被设计用于来自细胞培养物的细胞，所以不可能在紧凑组织例如神经索或卵巢上进行。 因此，我们适应LC3B标签初始协议，以能够标记来自陆地isopods的脂肪细胞的自噬膜，形成松散的组织，可以更容易被抗体穿透。 以下方案允许直接在动物中新鲜取样的陆生等足细胞中观察自噬泡。
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