Product Pathways - Apoptosis / Autophagy
LC3A (D50G8) XP® Rabbit mAb #4599
PhosphoSitePlus® protein, site, and accession data: LC3A
| Applications | Reactivity | Sensitivity | MW (kDa) | Isotype |
|---|---|---|---|---|
| W IP IHC-P IF-IC F | H M R (Mk) (Dg) | Endogenous | 14, 16 | Rabbit IgG |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
IHC-P=Immunohistochemistry (Paraffin)
IF-IC=Immunofluorescence (Immunocytochemistry)
F=Flow Cytometry
Reactivity Key:
H=Human
M=Mouse
R=Rat
Mk=Monkey
Dg=Dog
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Protocols
* Product-specific protocol.
Specificity / Sensitivity
LC3A (D50G8) XP® Rabbit mAb detects endogenous levels of total LC3A protein. This antibody may also react with LC3B.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human LC3A.
Western Blotting
Western blot analysis of extracts from HeLa and NIH/3T3 cells, untreated or chloroquine-treated (50 μM, overnight), using LC3A (D50G8) XP® Rabbit mAb.
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human glioblastoma multiforme using LC3A (D50G8) XP® Rabbit mAb.
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using LC3A (D50G8) XP® Rabbit mAb (left) orRabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded HeLa cell pellets, control (left) or choloroquine-treated (right), using LC3A (D50G8) XP® Rabbit mAb.
Flow Cytometry
Flow cytometric analysis of Chloroquine-treated HeLa cells, using LC3A (D50G8) XP® Rabbit mAb (blue) compared to Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (red).
IF-IC
Confocal immunofluorescent analysis of HeLa cells, untreated (left) or chloroquine-treated (right), using LC3A (D50G8) XP® Rabbit mAb (green). Actin filaments were labeled using DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (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). 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).
- 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
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
Companion Products
- 4108 LC3A/B Antibody
- 3868 LC3B (D11) XP® Rabbit mAb
- 2775 LC3B Antibody
- 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
- 8112 SignalStain® Antibody Diluent
- 8114 SignalStain® Boost IHC Detection Reagent (HRP, Rabbit)
This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.