自噬形成时，胞浆型LC3（即LC3-I）会酶解掉一小段多肽，转变为（自噬体）膜型（即LC3-II），因此可利用WB检测LC3-II/I比值的变化以评估自噬的强弱。此外自噬衔接物如sequestosome 1（SQSTM1，也称为p62）也可用于检测自噬，p62蛋白水平的多少与自噬强弱有着反比例关系。2.3荧光蛋白标记检测 2.3.1 GFP-LC3 ...
如下图:细胞转染mRFP-GFP-LC3病毒后给予氨基酸剥夺处理2小时后出现明显增强的自噬以及自噬流。 红色斑点是自噬溶酶体(mRFP),黄色斑点是自噬体(RFP+GFP). 通过不同颜色斑点的计数可以清晰看出自噬流的强弱,实时监测自噬发生过程,准确,清晰,直观! 参考文献
TagRFP-LC3 可识别从吞噬细胞到自噬体的自噬结构，在雷帕霉素处理的 MEFs 中DALGreen标记了61.8%的TagRFP-LC3点（自噬溶酶体阶段），其余的TagRFP-LC3点被认为是吞噬细胞和自噬体。DAPRed探针标记了65.7%的GFP-LC3点（吞噬体、自噬小体和自噬溶酶体）。其余的 GFP-LC3 点被认为是早期吞噬体。CQ（或 BafA1）...
目前文献对自噬过程进行观察和检测常用的策略和手段有:通过western blot检测LC3的剪切;通过电镜观测自噬体的形成;在荧光显微镜下采用GFP(-RFP)-LC3等融合蛋白来示踪自噬体形成以及降解。近几年对自噬流的研究日趋增多,针对于此我们汉恒生物科技(上海)有限公司自主研发了用于实时监测自噬(流)的mRFP-GFP-LC3腺病毒,mRFP...
Robust - Since autophagy is a constitutive cellular degradation process, the use of lysosomal inhibitors prevents the degradation of GFP-LC3, prolonging the signal and enabling quantification by flow cytometry. Low cost – stable cell lines allow repeated use o...
图3激光共聚焦下观察RFP-GFP-LC3稳定细胞株中自噬体的形成 (A)2h血清饥饿后,RFP-GFP-LC3稳定细胞株中自噬体的形成图(标尺50μm);(B)图A中绿色荧光点/红色荧光点的统计结果。**:P<0.01,与对照组相比;#:P<0.05,与饥饿组相比。 Fig.3ObservationofautophagyformationinRFP-GFP-LC3stablecelllineunderlaserscann...
The Premo™ Autophagy Tandem Sensor RFP-GFP-LC3B Kit combines the ability to monitor the various stages of autophagy (through LC3B protein localization) with the high transduction efficiency and minimal toxicity of BacMam 2.0 expression technology. To perform image-based analysis for autophagy, ...
吉满助力使用pGMLV-GFP-LC3 慢病毒(Genomeditech Co., Ltd.)感染CMEC细胞，使用荧光显微镜观察经过DOX和Irisin处理的CMEC细胞发现(图O-R)，Irisin可以促进CMEC细胞自噬的形成，并且可以部分挽救CMEC细胞中因DOX造成的自噬紊乱。案例二：N6-methyladenosine modification regulates ferroptosis through autophagy signaling pathway...
This probe is cleaved to produce GFP-LC3 and RFP-LC3螖G, the former of which is degraded by autophagy, and the latter of which is maintained in the cytoplasm to serve as an internal control. Autophagic activity can be simply and quantitatively estimated by determining the ratio of GFP and ...
GFP reached to 100%. The number of autophagic puncta significantly increased after starvationtreatment. Conclusion The RAW264.7 cell strain with stable expression of RFP-GFP-LC3 has been successfully constructed,which provides a reliable cellular platform for autophagy research.[Key words] autophagy;...