Cell Signaling Technology

Product Pathways - Autophagy Signaling

LC3 Control Cell Extracts #11972

extract   LC3   LC3A   LC3A/B   LC3B   lysate  

No. Size Price
11972S 100 µl ( 10 western blots ) ¥1,415.00 现货查询 购买询价
11972 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype

Species cross-reactivity is determined by western blot.

Applications Key:

Description

Total cell extracts from HeLa cells were untreated or treated with 50 μM chloroquine overnight. Supplied in SDS Sample Buffer. Boil for 2 minutes in the original tube, then load 10 μl per mini-gel lane.

Hela细胞总裂解液经 50μM 氯喹过夜处理或未经处理。加入SDS上样缓冲液。在原管中煮沸2分钟,微型凝胶孔上样10μl/孔。

Western Blotting

Western Blotting

Western blot analysis of LC3 Control Cell Extracts from HeLa cells, untreated (-) or chloroquine-treated (50 μM, overnight; +), using LC3B (D11) XP® Rabbit mAb #3868 (upper), LC3A/B Antibody #4108 (middle), or β-Tubulin (9F3) Rabbit mAb #2128 (lower).

Western blot检测来自Hela细胞的LC3对照细胞提取物,未处理(-)或氯喹处理(50μM,过夜,+),使用的抗体是LC3B (D11) XP® Rabbit mAb 兔单抗#3868(上图),LC3A/B Antibody #4108(中图),或β-Tubulin (9F3) Rabbit mAb 兔单抗#2128(下图)。

Applications

Western Blots: CST recommends boiling control cell extracts for two minutes prior to loading. Load 10 µl of each cell extract per lane.

Western Blots:CST建议加样前将细胞提取物煮沸2分钟。每孔加入10µl细胞样品。

Background

Autophagy is a catabolic process for the autophagosomic-lysosomal degradation of bulk cytoplasmic contents (1,2). Autophagy is generally activated by conditions of nutrient deprivation, but it has also been associated with a number of physiological processes including development, differentiation, neurodegenerative diseases, infection, and cancer (3). Autophagy marker Light Chain 3 (LC3) was originally identified as a subunit of microtubule-associated proteins 1A and 1B (termed MAP1LC3) (4) and subsequently found to contain similarity to the yeast protein Apg8/Aut7/Cvt5 critical for autophagy (5). Three human LC3 isoforms (LC3A, LC3B, and LC3C) undergo post-translational modifications during autophagy (6-9). Cleavage of LC3 at the carboxy terminus immediately following synthesis yields the cytosolic LC3-I form. During autophagy, LC3-I is converted to LC3-II through lipidation by a ubiquitin-like system involving Atg7 and Atg3 that allows for LC3 to become associated with autophagic vesicles (6-10). The presence of LC3 in autophagosomes and the conversion of LC3 to the lower migrating form, LC3-II, have been used as indicators of autophagy (11).

自噬是自噬溶酶体对其包裹的细胞质内含物进行降解的一种分解代谢过程(1,2)。自噬过程一般是在营养匮乏的条件下被激活,但是也与一些生理过程有关,如发育、分化、神经变性、感染和癌症(3)。自噬微管相关蛋白的轻链3(LC3)最初被认定为是微管相关蛋白1A和1B的一个亚基(被称为MAP1LC3)(4),随后发现该蛋白所包含的与酵母蛋白类似的Apg8/Aut7/Cvt5对于自噬起着至关重要的作用(5)。三种人类LC3亚型(LC3A、LC3B和LC3C)在自噬过程中翻译后被修饰(6-9)。随着合成后LC3羧基末端迅速裂解,产生了胞浆的LC3-I 型。在自噬过程中,通过一个涉及Atg7和Agt3的类泛素体系,LC3-I被脂质化为LC3-II,从而将LC3与自噬小泡联系起来(6-10)。自噬体中LC3的存在,及其向低迁移形式的LC3-II的转化被作为自噬发生的“指示器”(11)。

  1. Reggiori, F. and Klionsky, D.J. (2002) Eukaryot. Cell 1, 11-21.
  2. Codogno, P. and Meijer, A.J. (2005) Cell Death Differ. 12 Suppl 2, 1509-18.
  3. Levine, B. and Yuan, J. (2005) J. Clin. Invest. 115, 2679-88.
  4. Mann, S.S. and Hammarback, J.A. (1994) J. Biol. Chem. 269, 11492-97.
  5. Lang, T. et al. (1998) EMBO J. 17, 3597-607.
  6. Kabeya, Y. et al. (2000) EMBO J. 19, 5720-28.
  7. He, H. et al. (2003) J. Biol. Chem. 278, 29278-87.
  8. Tanida, I. et al. (2004) J. Biol. Chem. 279, 47704-10.
  9. Wu, J. et al. (2006) Biochem. Biophys. Res. Commun. 339, 437-42.
  10. Ichimura, Y. et al. (2000) Nature 408, 488-92.
  11. Kabeya, Y. et al. (2004) J. Cell Sci. 117, 2805-12.

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