Product Pathways - Apoptosis / Autophagy
LC3B Antibody #2775
| Applications | Reactivity | Sensitivity | MW (kDa) | Source |
|---|---|---|---|---|
| W IF-IC | H M R (Mk) (C) (B) (X) | Endogenous | 14, 16 | Rabbit |
Applications Key:
W=Western Blotting
IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:
H=Human
M=Mouse
R=Rat
Mk=Monkey
C=Chicken
B=Bovine
X=Xenopus
Species cross-reactivity is determined by Western blot.
Specificity / Sensitivity
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.
Source / Purification
Polyclonal antibodies are produced by immunizing rabbits with a synthetic peptide (KLH-coupled) 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.
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™ (fluorescent DNA dye).
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 has also been associated with a number of physiological processes including development, differentiation, neurodegenerative diseases, infection and cancer (3). Light chain 3 (LC3), which can serve as a marker for autophagy, 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). There are three human isoforms of LC3 - LC3A, LC3B, and LC3C - that undergo post-translational modifications during autophagy (6-9). LC3 is first cleaved at the carboxy terminus immediately following synthesis to yield a cytosolic form LC3-I. 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). Thus, the presence of LC3 in autophagosomes as well as the conversion of LC3 to the lower migrating form LC3-II have been used as indicators of autophagy.
- Reggiori, F. and Klionsky, D.J. (2002) Eukaryot. Cell 1, 11-21.
- Codogno, P. and Meijer, A.J. (2005) Cell Death Differ. 12 Suppl 2, 1509-1518.
- Levine, B. and Yuan, J. (2005) J. Clin. Invest. 115, 2679-2688.
- Mann, S.S. and Hammarback, J.A. (1994) J. Biol. Chem. 269, 11492-11497.
- Lang, T. et al. (1998) EMBO J. 17, 3597-3607.
- Kabeya, Y. et al. (2000) EMBO J. 19, 5720-5728.
- He, H. et al. (2003) J. Biol. Chem. 278, 29278-29287.
- Tanida, I. et al. (2004) J. Biol. Chem. 279, 47704-47710.
- Wu, J. et al. (2006) Biochem. Biophys. Res. Commun. 339, 437-442.
- Ichimura, Y. et al. (2000) Nature 408, 488-492.
- Kabeya, Y. et al. (2004) J. Cell Sci. 117, 2805-2812.
Application References
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This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.
