LC3B (MAP1LC3B) (N-term) (incl. pos. control) Mouse Monoclonal Antibody [Clone ID: 5F10]

Specifications

Product Data
Clone Name	5F10
Applications	IF, WB
Recommend Dilution	Immunocytochemistry: Use at 1-10 µg/ml (Paraformaldehyd/Methanol fixation) (See Ref.1) Immunoblotting: 0.5 µg/ml for HRPO/ECL detection Recommended blocking buffer: Casein/Tween 20 based blocking and blot incubation buffer. We strongly recommend to use PVDF membranes for Immunoblot analysis. Included Positive Control Cell Lysates: Positive Control: Enriched cell fraction (LC3 I and LC3II) from PC3 cells, Format: Lyophilized cell lysate from PC3 cells. Reconstitution: Restore by addition of 200 µl H2O. After complete solubilization add 200 µl 2x SDS-PAGE sample buffer, mix and incubate at 90°C for 5 min. Application: The positive control cell lysate is recommended for immunoblot applications. 20 µl of positive control cell lysate correspond to ca. 20.000 cells. Use 20 µl/lane (mini gel) for HRPO/ECL detection of the target proteins. Please NOTE: The lyophilized cell lysates contains SDS and are not recommended for applications with native proteins such as in immunoprecipitation. Storage: Aliquote reconstituted product and store frozen. Avoid repeated fereezing and thawing.
Reactivity	Canine, Hamster, Human, Mouse, Rat
Host	Mouse
Clonality	Monoclonal
Immunogen	Synthetic peptide - hemocyanin conjugated - derived from the N-terminus of LC3-B
Specificity	This antibody specifically recognizes both forms of endogenous LC3, the cytoplasmic LC3-I (18 kDa) as well as the lipidated form generated during autophagosome and autophagolysosome formation: LC3-II (16 kDa). Immunocytochemical staining of cells with LC3 antibody  Cat.-No AM20212PU-N (Clone 5F10) reveals the specific punctate distribution of endogenous LC3-II as a hallmark of autophagic activity.
Formulation	1ml PBS State: Purified State: Lyophilized purified IgG fraction Stabilizer: PEG and Sucrose Preservative: 0.09% Sodium Azide
Reconstitution Method	Restore with 1ml H2O (15 min, RT)
Purification	Subsequent Ultrafiltration and Size Exclusion Chromatography
Conjugation	Unconjugated
Storage Condition	Store lyophilized (preferably in a desiccator) at -20°C and reconstituted (aliquote and freeze in liquid nitrogen) at -80°C. Thaw aliquots at 37°C. Thawed aliquots may be stored at 4°C up to 3 months. Avoid repeated freezing and thawing.
Predicted Protein Size	18 kDa (LC3-I), 16 kDa (LC-II)
Gene Name	microtubule associated protein 1 light chain 3 beta
Database Link	Entrez Gene 64862 RatEntrez Gene 67443 MouseEntrez Gene 81631 Human UniProt ID Q9GZQ8
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.
Synonyms	MAP1LC3B, MAP1A/MAP1B, Map1lc3b, Map1alc3, Map1lc3
Reference Data

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