Product Details
| Alternative Name: | Microtubule-associated protein 1 light chain 3B, MAP1LC3B |
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| Clone: | 5F10 |
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| Host: | Mouse |
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| Isotype: | IgG1 |
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| Immunogen: | Synthetic peptide corresponding to the N-terminus of LC3B conjugated to hemocyanin. |
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| UniProt ID: | Q9GZQ8 |
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| Species reactivity: | Human, Mouse, Rat
Dog, Hamster
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| Specificity: | Recognizes both forms of endogenous LC3B. |
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| Applications: | ICC, WB
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| Recommended Dilutions/Conditions: | Immunocytochemistry (paraformaldehyde/methanol fixed cells; 1-10 µg/ml)
Western Blot (0.5µg/ml for HRPO/ECL detection; recommended blocking buffer CPPT: 10mM TRIS-HCl, pH 7.4, 0.5% (w/v) casein, 1% (w/v) PEG 4000, 1% (w/v) polyvinylpyrrolidone, 0.1% (v/v) Tween 20, 150mM sodium chloride)
We strongly recommend to use PVDF membranes for immunoblot analysis.
Suggested dilutions/conditions may not be available for all applications.
Optimal conditions must be determined individually for each application. |
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| Application Notes: | Detects a band of ~18kDa (LC3B-I; cytoplasmic form) and a band of ~16kDa (LC3B-II; lipidated form) by Western blot. |
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| Positive Control: | Included. (Prod. No. ALX-840-038) |
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| Purity Detail: | Purified from serum-free cell culture supernatant by subsequent ultrafiltration and size exclusion chromatography. |
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| Formulation: | Lyophilized from 1ml PBS containing PEG, sucrose, and 0.09% sodium azide. |
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| Reconstitution: | Reconstitute with 1ml water (15 minutes at room temperature). |
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| Use/Stability: | Stable at -80°C up to 1 year, at 4°C up to 3 months. |
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| Handling: | Avoid freeze/thaw cycles. After reconstitution, prepare aliquots and freeze in liquid nitrogen. |
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| Shipping: | Blue Ice |
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| Long Term Storage: | -20°C |
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| Scientific Background: | Autophagy is an alternative process of proteasomal degradation for some long-lived proteins or organelles. Alterations in the autophagic-lysosomal compartment have been linked to neuronal death in many neurodegenerative disorders as well as in transmissible neuronal pathologies (prion diseases). Genetic studies in yeast have shown that Autophagy-defective Gene-8 (Atg-8) represents a specific marker for autophagy. Among the four families of mammalian Atg8-related proteins only LC3 (Microtubule-associated Protein1 Light Chain 3) is expressed at sufficient high levels and efficiently recruited to autophagic vesicles in cells and tissues. During autophagy the cytoplasmic form, LC3-I is processed and recruited to autophagosomes, where LC3-II is generated by site specific proteolysis near to the C-terminus. Autophagic vacuoles have been also reported frequently in cardiomyopathies or muscle cells exposed to different experimental settings. |
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| Regulatory Status: | RUO - Research Use Only |
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Figure 1: Endogenous LC3 punctae detected using MAb to LC3B (5F10) (Prod. No. ALX-803-080)Method: The majority of LC3 was diffusely localized in unstimulated COS-7 cells, wheres punctated signals of LC3 increase after induction of autophagy by vinblastin stimulation for 2h. Cells were fixed with paraformaldehyde followed by methanol treatment. Cells were permeabilized with 0.3% Triton X-100. (Image courtesy of I. Ciechomska and A. Tolkovsky, University of Cambridge, UK.)
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Product Literature References
Squalene-Based Nano-Assemblies Improve the Pro-Autophagic Activity of Trehalose: G. Frapporti, et al.; Pharmaceutics
14, 862 (2022),
Abstract;
The unfolded protein response transducer IRE1α promotes reticulophagy in podocytes: J.R.N. Betancourt, et al.; Biochim. Biophys. Acta Mol. Basis Dis.
1868, 166391 (2022),
Abstract;
DRP1 haploinsufficiency attenuates cardiac ischemia/reperfusion injuries: L. Bouche, et al.; PLoS One
16, 248554 (2021),
Abstract;
Effect of Klotho on autophagy clearance in tacrolimus-induced renal injury: S.W. Lim, et al.; FASEB J.
33, 2694 (2019),
Abstract;
Autophagy regulates DUOX1 localization and superoxide production in airway epithelial cells during chronic IL-13 stimulation: J.D. Dickinson, et al.; Redox Biol.
14, 272 (2018),
Abstract;
Full Text
Laparoscopic Sleeve Gastrectomy Improves Nonalcoholic Fatty Liver Disease-Related Liver Damage in Adolescents by Reshaping Cellular Interactions and Hepatic Adipocytokine Production: V. Nobili, et al.; J. Pediatr.
194, 100 (2018),
Abstract;
Ginseng extract reduces tacrolimus-induced oxidative stress by modulating autophagy in pancreatic beta cells: S.W. Lim, et al.; Lab. Invest.
97, 1271 (2017),
Abstract;
Effect of Exendin-4 on Autophagy Clearance in Beta Cell of Rats with Tacrolimus-induced Diabetes Mellitus: S.W. Lim, et al.; Sci. Rep.
6, 29921 (2016),
Application(s): Primary antibodies used for immunoblot analysis,
Abstract;
Full Text
Nordihydroguaiaretic acid (NDGA) and α-mangostin inhibit the growth of Mycobacterium tuberculosis by inducing autophagy: S. Guzman-Beltran, et al.; Int. Immunopharmacol.
31, 149 (2016),
Application(s): LC3 immunofluorescent staining,
Abstract;
AD-linked, toxic NH2 human tau affects the quality control of mitochondria in neurons: G. Amadoro, et al.; Neurobiol. Dis.
62, 489 (2014),
Abstract;
Phosphorylation of NBR1 by GSK3 modulates protein aggregation: N.S. Nicot, et al.; Autophagy
10, 1036 (2014),
Abstract;
Full Text
Autophagy facilitates organelle clearance during differentiation of human erythroblasts: evidence for a role for ATG4 paralogs during autophagosome maturation: V.M. Betin, et al.; Autophagy
9, 881 (2013),
Abstract;
Full Text
The nuclear cofactor DOR regulates autophagy in mammalian and Drosophila cells: C. Mauvezin, et al.; EMBO Rep.
11, 37 (2010),
Abstract;
Full Text
Discovery of Atg5/Atg7-independent alternative macroautophagy: Y. Nishida, et al.; Nature
461, 654 (2009),
Abstract;
Early endosomes and endosomal coatomer are required for autophagy: M. Razi, et al.; J. Cell. Biol.
185, 305 (2009),
Abstract;
