Revision 1

#8666Store at -20C

1 Kit

(3 x 20 microliters)

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
Atg12 Antibody 2010 20 µl 16, 55 kDa Rabbit 
LC3A (D50G8) XP® Rabbit mAb 4599 20 µl 14, 16 kDa Rabbit IgG
LC3B (D11) XP® Rabbit mAb 3868 20 µl 14, 16 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The Autophagosome Marker Antibody Sampler Kit provides an economical means to investigate the accumulation of autophagosomes within the cell. The kit contains enough primary and secondary antibodies to perform two western blots per primary antibody.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

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, neurodegeneration, infection, and cancer (3). The molecular machinery of autophagy was largely discovered in yeast and referred to as autophagy-related (Atg) genes. Formation of the autophagosome involves a ubiquitin-like conjugation system in which Atg12 is covalently bound to Atg5 and targeted to autophagosome vesicles (4-6). This conjugation reaction is mediated by the ubiquitin E1-like enzyme Atg7 and the E2-like enzyme Atg10 (7,8). Autophagy marker Light Chain 3 (LC3) was originally identified as a subunit of microtubule-associated proteins 1A and 1B (termed MAP1LC3) (9) and subsequently found to contain similarity to the yeast protein Apg8/Aut7/Cvt5 that is critical for autophagy (10). Three human LC3 isoforms (LC3A, LC3B, and LC3C) undergo post-translational modifications during autophagy (11-14). 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 (11-15). 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 (16).

  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. Mizushima, N. et al. (1998) J Biol Chem 273, 33889-92.
  5. Mizushima, N. et al. (1998) Nature 395, 395-8.
  6. Suzuki, K. et al. (2001) EMBO J 20, 5971-81.
  7. Tanida, I. et al. (1999) Mol Biol Cell 10, 1367-79.
  8. Shintani, T. et al. (1999) EMBO J 18, 5234-41.
  9. Mann, S.S. and Hammarback, J.A. (1994) J Biol Chem 269, 11492-7.
  10. Lang, T. et al. (1998) EMBO J 17, 3597-607.
  11. Kabeya, Y. et al. (2000) EMBO J 19, 5720-8.
  12. Wu, J. et al. (2006) Biochem Biophys Res Commun 339, 437-42.
  13. He, H. et al. (2003) J Biol Chem 278, 29278-87.
  14. Tanida, I. et al. (2004) J Biol Chem 279, 47704-10.
  15. Ichimura, Y. et al. (2000) Nature 408, 488-92.
  16. Kabeya, Y. et al. (2004) J Cell Sci 117, 2805-12.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    Limited Uses

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