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LC3B monoclonal antibody (5F10)

 
ALX-803-080-C100 100 µg 528.00 USD
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Product Specification

Alternative Name:Microtubule-associated protein 1 light chain 3B, MAP1LC3B
 
Clone:5F10
 
Host:Mouse
 
Isotype:IgG1
 
Immunogen:Synthetic peptide corresponding to the N-terminus of LC3B conjugated to hemocyanin.
 
UniProt ID:Q9GZQ8
 
Species reactivity:Human, Mouse, Rat
Dog, Hamster
 
Specificity:Recognizes both forms of endogenous LC3B.
 
Applications:ICC, WB
 
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.
 
Application Notes:Detects a band of ~18kDa (LC3B-I; cytoplasmic form) and a band of ~16kDa (LC3B-II; lipidated form) by Western blot.
 
Positive Control:Included. (Prod. No. ALX-840-038)
 
Purity Detail:Purified from serum-free cell culture supernatant by subsequent ultrafiltration and size exclusion chromatography.
 
Formulation:Lyophilized from 1ml PBS containing PEG, sucrose, and 0.09% sodium azide.
 
Reconstitution:Reconstitute with 1ml water (15 minutes at room temperature).
 
Use/Stability:Stable at -80°C up to 1 year, at 4°C up to 3 months.
 
Handling:Avoid freeze/thaw cycles. After reconstitution, prepare aliquots and freeze in liquid nitrogen.
 
Shipping:Shipped on Blue Ice
 
Long Term Storage:-20°C
 
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.
 
803-080
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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803-080

Product Literature References

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;
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;

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