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PDP - Template Name: Antibody Sampler Kit
PDP - Template ID: *******4a3ef3a
  1. Products /
  2. Primary Antibodies /
  3. Autophagy Vesicle Elongation (LC3 Conjugation) Antibody Sampler Kit

Autophagy Vesicle Elongation (LC3 Conjugation) Antibody Sampler Kit #19848

    Product Information

    Kit Usage Information

    Protocols

    Product 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.

    Specificity / Sensitivity

    LC3A/B (D3U4C) XP® Rabbit mAb recognizes endogenous levels of total LC3A and LC3B proteins. Atg7 (D12B11) Rabbit mAb recognizes endogenous levels of total Atg7 protein. Atg4B (D1G2R) Rabbit mAb recognizes endogenous levels of total Atg4B protein. Atg4A (D62C10) Rabbit mAb recognizes endogenous levels of total Atg4A protein and recognizes unidentified bands within the molecular weight range of 48-60 kDa, which decrease with silencing of Atg4A expression. GABARAP (E1J4E) Rabbit mAb recognizes endogenous levels of total GABARAP protein, but does not cross-react with other GABARAP family members. Atg3 Antibody detects endogenous levels of total Atg3 protein.

    Source / Purification

    Monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu44 of human LC3B protein (conserved in LC3A), a synthetic peptide corresponding to residues near the amino terminus of human Atg7 protein, a synthetic peptide corresponding to residues surrounding Gln100 of human Atg4B protein, a synthetic peptide corresponding to residues near the carboxy terminus of human Atg4A protein, a synthetic peptide corresponding to residues surrounding Arg40 of human GABARAP protein. Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of Atg3. Antibodies are purified by protein A and peptide affinity chromatography.

    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.

    Database Links

    UniProt ID: Q9Y4P1 , Q9H492 , Q9GZQ8 , O95166 , Q8WYN0 , Q9NT62 , O95352


    Entrez-Gene Id: 23192 , 84557 , 81631 , 11337 , 115201 , 64422 , 10533


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    Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

    For Research Use Only. Not For Use In Diagnostic Procedures.
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