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useful for the research of a role in apoptosis, mitochondrial morphology and autophagy

Anti LC3 (Clone: LC3-1703) monoclonal antibody

Background

LC3B is one of the mammalian Atg8 homologs and widely used as an autophagosome marker. Immediately after synthesis, LC3 is processed by Atg4 and becomes LC3-I. Upon induction of autophagy, the C-terminal glycine of LC3-I is conjugated to phosphatidylethanolamine, resulting in formation of membrane-bound LC3-II. Most LC3-II is thought be present on autophagosome membrane. The autophagosome subsequently fuses with a lysosome, where inside materials, including LC3-II, are degraded. The expression level of LC3-II generally correlates with the number of autophagosome.

Application

<Immunofluorescence microscopy analysis of mouse embryonic fibroblasts (MEFs)>
Anti LC3 antibody
MEFs were cultured in regular DMEM supplemented with 10% FBS (left) or DMEM without amino acids (right) for 1 hr. After fixation with 4% paraformaldehyde for 10 min at room temperature, they were permeabilized with 50 µg/ml digitonin for 5 min. Cells were then subjected to immunofluorescence microscopy using #CAC-CTB-LC3-2-IC anti-LC3 antibody at 1:100 dilution. Scale bar, 20 µm.

< Immuno-electron microscopy analysis of mouse embryonic fibroblasts (MEFs)>
Anti LC3 antibody
MEFs were cultured in DMEM without amino acids for 2 hr. After fixation with 4% paraformaldehyde for 2 hr at room temperature, they were permeabilized with liquid nitrogen. Immuno-electron microscopy analysis (pre-embedding method) of endogenous LC3 was performed using  #CAC-CTB-LC3-2-IC anti-LC3 antibody at 1:10 dilution. The gold labeling was intensified by using a silver enhancement kit (HQ silver enhancement kit, Nanoprobes, NY). Scale bar, 500 nm.

Applying Campaign Price (from July 1st - November 30th, 2017)

⇒ Autophagy antibody products 30% discount campaign

Product List

Product Name Cat# Quantity Price

Anti LC3

CAC-CTB-LC3-2-IC 50UG

¥ 42,000
$ 800
€ 600

Citation
  • (1)Small GTPase Rab1B is associated with ATG9A vesicles and regulates autophagosome formation.
    Kakuta S, Yamaguchi J, Suzuki C, Sasaki M, Kazuno S, Uchiyama Y.
    FASEB J. 2017 May 18. pii: fj.201601052R. PMID: 28522593
  • (2)Cholesterol and ORP1L-mediated ER contact sites control autophagosome transport and fusion with the endocytic pathway.
    Wijdeven RH, Janssen H, Nahidiazar L, Janssen L, Jalink K, Berlin I, Neefjes J.
    Nat Commun. 2016 Jun 10;7:11808. PMID: 27283760
  • (3)Helicobacter pylori VacA induces apoptosis by accumulation of connexin 43 in autophagic vesicles via a Rac1/ERK-dependent pathway.
    Yahiro K, Akazawa Y, Nakano M, Suzuki H, Hisatune J, Isomoto H, Sap J, Noda M, Moss J, Hirayama T.
    Cell Death Discov. 2015 Sep 28;1:15035. PMID: 27551466
  • (4)The versatile electron microscope: an ultrastructural overview of autophagy.
    Biazik J, Vihinen H, Anwar T, Jokitalo E, Eskelinen EL
    Methods. 2015 Mar;75:44-53. PMID: 25433244
  • (5)Ultrastructural analysis of autophagosome organization using mammalian autophagy-deficient cells.
    Kishi-Itakura C, Koyama-Honda I, Itakura E, Mizushima N.
    J Cell Sci. 2014 Sep 15;127(Pt 18):4089-102. PMID: 25052093
  • (6)DAP1, a negative regulator of autophagy, controls SubAB-mediated apoptosis and autophagy.
    Yahiro K, Tsutsuki H, Ogura K, Nagasawa S, Moss J, Noda M.
    Infect Immun. 2014 Nov;82(11):4899-908. PMID: 25183729
  • (7)The hairpin-type tail-anchored SNARE syntaxin 17 targets to autophagosomes for fusion with endosomes/lysosomes.
    Itakura E, Kishi-Itakura C, Mizushima N.
    Cell. 2012 Dec 7;151(6):1256-69. PMID: 23217709
  • (8)Low-density lipoprotein receptor-related protein-1 (LRP1) mediates autophagy and apoptosis caused by Helicobacter pylori VacA.
    Yahiro K, Satoh M, Nakano M, Hisatsune J, Isomoto H, Sap J, Suzuki H, Nomura F, Noda M, Moss J, Hirayama T.
    J Biol Chem. 2012 Sep 7;287(37):31104-15. PMID: 22822085
  • (9)Mammalian Atg2 proteins are essential for autophagosome formation and important for regulation of size and distribution of lipid droplets.
    Velikkakath AK, Nishimura T, Oita E, Ishihara N, Mizushima N.
    Mol Biol Cell. 2012 Mar;23(5):896-909. PMID: 22219374
  • (10)De novo mutations in the autophagy gene WDR45 cause static encephalopathy of childhood with neurodegeneration in adulthood.
    Saitsu H, Nishimura T, Muramatsu K, Kodera H, Kumada S, Sugai K, Kasai-Yoshida E, Sawaura N, Nishida H, Hoshino A, Ryujin F, Yoshioka S, Nishiyama K, Kondo Y, Tsurusaki Y, Nakashima M, Miyake N, Arakawa H, Kato M, Mizushima N, Matsumoto N.
    Nat Genet. 2013 Apr;45(4):445-9, 449e1. doi: 10.1038/ng.2562. Epub 2013 Feb 24 PMID: 23435086

References
  • (1)Methods in mammalian autophagy research.
    Mizushima N, Yoshimori T, Levine B.
    Cell. 2010 Feb 5;140(3):313-26. PMID: 20144757
  • (2)How to interpret LC3 immunoblotting.
    Mizushima N, Yoshimori T.
    Autophagy. 2007 Nov-Dec;3(6):542-5. PMID: 17611390
  • (3)LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing.
    Kabeya Y, Mizushima N, Ueno T, Yamamoto A, Kirisako T, Noda T, Kominami E, Ohsumi Y, Yoshimori T.
    EMBO J. 2000 Nov 1;19(21):5720-8. PMID: 11060023

To be used for research only. DO NOT use for human gene therapy or clinical diagnosis.