FastScan™ Total LC3B ELISA Kit #35172
Important Ordering Details
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Supporting Data
| REACTIVITY | H M R Mk |
Species Cross-Reactivity Key:
- H-Human
- M-Mouse
- R-Rat
- Mk-Monkey
Product Information
Product Description
The FastScan™ Total LC3B ELISA Kit is a sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of LC3B. To perform the assay, sample is incubated with a capture antibody conjugated with a proprietary tag and a second detection antibody linked to HRP, forming a sandwich with LC3B in solution. This entire complex is immobilized to the plate via an anti-tag antibody. The wells are then washed to remove unbound material. TMB is then added. The magnitude of observed signal is proportional to the quantity of LC3B.
*Antibodies in kit are custom formulations specific to kit.
IMPORTANT: This FastScan™ ELISA Kit requires 4 washes at Step 6 of the protocol.
Protocol
Specificity / Sensitivity
The FastScan™ Total LC3B ELISA Kit detects endogenous levels of LC3B as shown in Figure 1. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
Species Reactivity:
Human, Mouse, Rat, Monkey
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 posttranslational 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, 1509-18.
- Levine, B. and Yuan, J. (2005) J. Clin. Invest. 115, 2679-88.
- Mann, S.S. and Hammarback, J.A. (1994) J. Biol. Chem. 269, 11492-97.
- Lang, T. et al. (1998) EMBO J. 17, 3597-607.
- Kabeya, Y. et al. (2000) EMBO J. 19, 5720-28.
- He, H. et al. (2003) J. Biol. Chem. 278, 29278-87.
- Tanida, I. et al. (2004) J. Biol. Chem. 279, 47704-10.
- Wu, J. et al. (2006) Biochem. Biophys. Res. Commun. 339, 437-42.
- Ichimura, Y. et al. (2000) Nature 408, 488-92.
- Kabeya, Y. et al. (2004) J. Cell Sci. 117, 2805-12.
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