Revision 2

#19848Store at -20C

1 Kit

(6 x 20 microliters)

Cell Signaling Technology

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

Support: 877-678-TECH (8324)

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For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
LC3A/B (D3U4C) XP® Rabbit mAb 12741 20 µl 14, 16 kDa Rabbit IgG
Atg7 (D12B11) Rabbit mAb 8558 20 µl 78 kDa Rabbit IgG
Atg4B (D1G2R) Rabbit mAb 13507 20 µl 48 kDa Rabbit IgG
Atg4A (D62C10) Rabbit mAb 7613 20 µl 48-60 kDa Rabbit IgG
GABARAP (E1J4E) Rabbit mAb 13733 20 µl 14, 16 kDa Rabbit IgG
Atg3 Antibody 3415 20 µl 40 kDa Rabbit 
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 Autophagy Vesicle Elongation (LC3 Conjugation) Antibody Sampler Kit provides an economical means of detecting target proteins related to autophagy vesicle elongation pathway. The kit contains enough antibody to perform two western blots per primary.

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 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 post-translational modifications during autophagy (6-8). 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 (6-9). 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 (10). Numerous mammalian counterparts to yeast Atg proteins have been described, including three Atg8 proteins (GATE-16, GABARAP, and LC3) and four Atg4 homologs (Atg4A/autophagin-2, Atg4B/autophagin-1, Atg4C/autophagin-3, and Atg4D/autophagin-4) (10-12). The cysteine protease Atg4 is pivotal to autophagosome membrane generation and regulation (13). GABAA receptor associated protein (GABARAP) is an Atg8 family protein with a key role in autophagy, which was originally discovered as a protein associated with the GABAA receptor regulating receptor trafficking to the plasma membrane (14). Processing of GABARAP involves cleavage by Atg4 family members (15,16) followed by conjugation by the E1 and E2 like enzymes Atg7 and Atg3 (17,18).

  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. Mann, S.S. and Hammarback, J.A. (1994) J Biol Chem 269, 11492-7.
  5. Lang, T. et al. (1998) EMBO J 17, 3597-607.
  6. He, H. et al. (2003) J Biol Chem 278, 29278-87.
  7. Tanida, I. et al. (2004) J Biol Chem 279, 47704-10.
  8. Wu, J. et al. (2006) Biochem Biophys Res Commun 339, 437-42.
  9. Ichimura, Y. et al. (2000) Nature 408, 488-92.
  10. Kabeya, Y. et al. (2004) J Cell Sci 117, 2805-12.
  11. Kabeya, Y. et al. (2000) EMBO J 19, 5720-8.
  12. Mariño, G. et al. (2003) J Biol Chem 278, 3671-8.
  13. Sou, Y.S. et al. (2008) Mol Biol Cell 19, 4762-75.
  14. Wang, H. et al. (1999) Nature 397, 69-72.
  15. Tanida, I. et al. (2004) J Biol Chem 279, 36268-76.
  16. Hemelaar, J. et al. (2003) J Biol Chem 278, 51841-50.
  17. Tanida, I. et al. (2001) J Biol Chem 276, 1701-6.
  18. Tanida, I. et al. (2002) J Biol Chem 277, 13739-44.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    XP is a registered trademark of Cell Signaling Technology, Inc.
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