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11972
LC3 Control Cell Extracts
Experimental Controls

LC3 Control Cell Extracts #11972

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  1. WB

Western blot analysis of LC3 Control Cell Extracts from HeLa cells, untreated (-) or chloroquine-treated (50 μM, overnight; +), using LC3B (D11) XP® Rabbit mAb #3868 (upper), LC3A/B Antibody #4108 (middle), or β-Tubulin (9F3) Rabbit mAb #2128 (lower).

Product Usage Information

Boil for 3 minutes prior to use. Load 10 µl of untreated and chloroquine treated LC3 Control Cell Extracts per lane.

Storage:

Supplied in SDS Sample Buffer: 62.5 mM Tris- HCl (pH 6.8 at 25°C), 2% w/v SDS, 10% glycerol, 50 mM DTT, 0.01% w/v bromophenol blue or phenol red.Store at –20°C, or at –80°C for long-term storage.

Product Description

LC3 Control Cell Extracts (HeLa Untreated): Total cell extracts from HeLa cells serve as a negative control. Supplied in SDS sample buffer.

LC3 Control Cell Extracts (HeLa +Chloroquine): Total cell extracts from HeLa cells treated with 50 μM chloroquine overnight serve as a positive control.

This lysate pair is produced as a control for western blotting of LC3A and LC3B. LC3C cannot be detected in these lysates.

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 (6-9). 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).

  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, 11492-97.
  5. Lang, T. et al. (1998) EMBO J. 17, 3597-607.
  6. Kabeya, Y. et al. (2000) EMBO J. 19, 5720-28.
  7. He, H. et al. (2003) J. Biol. Chem. 278, 29278-87.
  8. Tanida, I. et al. (2004) J. Biol. Chem. 279, 47704-10.
  9. Wu, J. et al. (2006) Biochem. Biophys. Res. Commun. 339, 437-42.
  10. Ichimura, Y. et al. (2000) Nature 408, 488-92.
  11. Kabeya, Y. et al. (2004) J. Cell Sci. 117, 2805-12.
For Research Use Only. Not For Use In Diagnostic Procedures.

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To Purchase # 11972S
Product # Size Price
11972S
100 µl  (10 western blots) $ 118