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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