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Product Name
LC3 antibody
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Description
LC3 Rabbit Polyclonal antibody. Positive IF detected in Starvation treated HEK-293 cells. Positive WB detected in Chloroquine treated HeLa cells, Chloroquine treated HEK-293 cells, human brain tissue, mouse brain tissue, rat brain tissue. Observed molecular weight by Western-blot: 16 kDa
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Tested applications
ELISA, IF, WB
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Species reactivity
Human,Mouse,Rat,Zebrafish; other species not tested.
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Alternative names
LC3 antibody; LC3A antibody; LC3A/B antibody; MAP1A/MAP1B LC3 A antibody; MAP1A/MAP1B light chain 3 A antibody; MAP1ALC3 antibody; MAP1BLC3 antibody; MAP1LC3A antibody
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Isotype
Rabbit IgG
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Preparation
This antibody was obtained by immunization of LC3 recombinant protein (Accession Number: XM_047440559). Purification method: Antigen affinity purified.
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Clonality
Polyclonal
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Formulation
PBS with 0.1% sodium azide and 50% glycerol pH 7.3.
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Storage instructions
Store at -20℃. DO NOT ALIQUOT
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Applications
Recommended Dilution:
WB: 1:500-1:5000
IF: 1:20-1:200
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Validations
Chloroquine treated HeLa cells were subjected to SDS PAGE followed by western blot with Catalog No:112163 (LC3 Antibody) at dilution of 1:1000
Immunofluorescent analysis of starvation treated HEK-293 cells using Catalog No:112163 (LC3 Antibody) at dilution of 1:50 and Alexa Fluor 488-congugated AffiniPure Goat Anti-Rabbit IgG(H+L). Cells were fixed with ethanol at -20 °C. 20 mM chloroquine was used to block the autophagy flux.
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Background
LC3, also known as microtubule-associated protein 1 light chain 3 (MAP1LC3), is a mammalian homolog of the yeast Atg8p (autophagy-related 8 protein) that is involved in autophagy process. Three isoforms of LC3 exist as LC3A,LC3B, and LC3C. LC3 is first synthesized as pro-LC3, and undergoes C-terminal cleavage to yield a cytosolic precursor known as LC3-I. During autophagy LC3-I is subsequently conjugated to PE lipid as lipidated form: LC3-II. The lipidated form migrates faster on SDS-PAGE gel. The increased the level of LC3-II has been widely used as autophagy marker. This antibody recognizes both LC3-I and LC3-II.
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References
- Kessel D, Oleinick NL. Initiation of autophagy by photodynamic therapy. Methods in enzymology. 453:1-16. 2009.
- Kessel D, Arroyo AS. Apoptotic and autophagic responses to Bcl-2 inhibition and photodamage. Photochemical & photobiological sciences : Official journal of the European Photochemistry Association and the European Society for Photobiology. 6(12):1290-5. 2007.
- Reiners JJ, Agostinis P, Berg K, Oleinick NL, Kessel D. Assessing autophagy in the context of photodynamic therapy. Autophagy. 6(1):7-18. 2010.
- Mariño G, Fernández AF, Cabrera S. Autophagy is essential for mouse sense of balance. The Journal of clinical investigation. 120(7):2331-44. 2010.
- Jin Y, Chen C, Meng L. A CE-LIF method to monitor autophagy by directly detecting LC3 proteins in HeLa cells. The Analyst. 137(23):5571-5. 2012.
- Niso-Santano M, Malik SA, Pietrocola F. Unsaturated fatty acids induce non-canonical autophagy. The EMBO journal. 34(8):1025-41. 2015.
- Liu M, Xu L, Yin L. Potent effects of dioscin against obesity in mice. Scientific reports. 5:7973. 2015.
- Zheng Q, Wang Q, Wang S, Wu J, Gao Q, Liu W. Thiopeptide Antibiotics Exhibit a Dual Mode of Action against Intracellular Pathogens by Affecting Both Host and Microbe. Chemistry & biology. 22(8):1002-7. 2015.
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