LC3B Antibody

2775:

Flow , Immunofluorescence* , Western Blotting

* Product-specific protocol.

LC3B detects endogenous levels of total LC3B protein. Cross-reactivity may exist with other LC3 isoforms. Stronger reactivity is observed with the type II form of LC3B.

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of LC3B. Antibodies were purified by peptide affinity chromatography.

Western Blotting

Western blot analysis of extracts from HeLa cells, mock transfected or transfected with rat LC3B, and from HT-1080 and A20 cells, untreated or chloroquine-treated (50 μM, overnight), using LC3B Antibody.

Flow Cytometry

Flow cytometric analysis of Hela cells using LC3B Antibody (blue) compared to a nonspecific negative control antibody (red).

IF-IC

Confocal immunofluorescent analysis of HeLa cells, untreated (left) or chloroquine-treated (right), using LC3B Antibody (green). Actin filaments have been labeled with Alexa Fluor^® 555 phalloidin (red). Blue pseudocolor = DRAQ5^® #4084(fluorescent DNA dye).

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 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).

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, 1509-1518.
3. Levine, B. and Yuan, J. (2005) J. Clin. Invest. 115, 2679-2688.
4. Mann, S.S. and Hammarback, J.A. (1994) J. Biol. Chem. 269, 11492-11497.
5. Lang, T. et al. (1998) EMBO J. 17, 3597-3607.
6. Kabeya, Y. et al. (2000) EMBO J. 19, 5720-5728.
7. He, H. et al. (2003) J. Biol. Chem. 278, 29278-29287.
8. Tanida, I. et al. (2004) J. Biol. Chem. 279, 47704-47710.
9. Wu, J. et al. (2006) Biochem. Biophys. Res. Commun. 339, 437-442.
10. Ichimura, Y. et al. (2000) Nature 408, 488-492.
11. Kabeya, Y. et al. (2004) J. Cell Sci. 117, 2805-2812.

• Li, C. et al. (2009) J Mol Cell Biol , . Applications: Western Blotting
• Chen, Y.F. et al. (2009) Genes Dev 23, 1183-94. Applications: Western Blotting
• Balakumaran, B.S. et al. (2009) Cancer Res 69, 7803-10. Applications: Western Blotting
• Mabuchi, S. et al. (2009) Clin Cancer Res 15, 5404-13. Applications: Western Blotting
• Deuretzbacher, A. et al. (2009) J Immunol 183, 5847-60. Applications: IF-IC (In Cells) Western Blotting
• Roca, H. et al. (2009) J Biol Chem 284, 34342-54. Applications: Western Blotting
• Alexander, A. et al. (2010) Proc Natl Acad Sci U S A 107, 4153-8. Applications: Western Blotting
• Wen, H. et al. (2011) Nat Immunol 12, 408-15. Applications: IF-IC (In Cells)
• Tong, Y. et al. (2012) Mol Neurodegener 7, 2. Applications: Western Blotting

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