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LC3B monoclonal antibody (5F10)



ALX-803-080-C100	100 µg	539.00 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.

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

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;