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LC3B monoclonal antibody (2G6)

 
ALX-803-081-C100 100 µg 528.00 USD
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Product Specification

Alternative Name:Microtubule-associated protein 1 light chain 3B, MAP1LC3B
 
Clone:2G6
 
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
Hamster, Monkey
 
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:Thiophilic adsorption and size exclusion chromatography purified.
 
Formulation:Liquid. In PBS containing 50% glycerol, 0.09% sodium azide, PEG and sucrose.
 
Use/Stability:Stable at -20°C up to 1 year.
 
Handling:Avoid freeze/thaw cycles.
 
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.
 
803-081
Figure: Western blot analysis of endogenous LC3 using MAb to LC3B (Prod. No. ALX-803-081).Method: Whole cell lysates of untreated  tumor cells were applied to SDS-PAGE and transferred to a PVDF membrane. The immunoblot was probed with Monoclonal Antibody to LC3 (2G6) (Prod. No. ALX-803-081)  (0.5 µg/ ml) for 1h at RT and developed by ECL (exposure time: 30 sec).

Lane 1: HeLa Lane 2: HepG2 Lane 3: HEK 293 Lane 4: SH-SY5Y Lane 5: MDCK Lane 6: PC12Lane 7: CMT Lane 8: Neuro2A Lane 9: NIH - 3T

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Product Literature References

Autophagy controls the pathogenicity of OPA1 mutations in dominant optic atrophy: M.S. Kane, et al.; J. Cell. Mol. Med. (2017), Abstract; Full Text

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