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    Anti-LC3B Rabbit mAb

    Catalog number :AT0763
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    Background: 
    Autophagy is a catabolic process for the autophagosomic-lysosomal degradation of bulk cytoplasmic contents . Autophagy is generally activated by conditions of nutrient deprivation, but it has also been associated with a number of physiological processes including development, differentiation, neurodegenerative diseases, infection, and cancer. Autophagy marker Light Chain 3 (LC3) was originally identified as a subunit of microtubule-associated proteins 1A and 1B (termed MAP1LC3) and subsequently found to contain similarity to the yeast protein Apg8/Aut7/Cvt5 critical for autophagy. Three human LC3 isoforms (LC3A, LC3B, and LC3C) undergo post-translational modifications during autophagy. Cleavage of LC3 at the carboxy terminus immediately following synthesis yields the cytosolic LC3-I form. During autophagy, LC3-I is converted to LC3-II through lipidation by a ubiquitin-like system involving Atg7 and Atg3 that allows for LC3 to become associated with autophagic vesicles. The presence of LC3 in autophagosomes and the conversion of LC3 to the lower migrating form, LC3-II, have been used as indicators of autophagy.
     
    UniProt ID: Q9GZQ8
    Overview
    Reactivity
    Human, Mouse, Rat, Pig
    Tested applications
    WB : 1/500-1/2000. 
    ICC/IF : 1/50-1/200.
    IP : 1/50
    Optimal dilutions/concentrations should be determined by the end user. 
    Specificity
    This antibody recognizes endogenous levels of total LC3B protein.
    Properties
    Immunogen
    a synthetic peptide corresponding to residues near the amino terminus of LC3B.
    Clonality
    Monoclonal, clone number: 4C13
    Isotype
    Rabbit IgG
    Form
    Liquid, 100 μl,1mg/ml, PBS (pH 7.2) and 40% Glycerol,0.02% Sodium Azide
    Storage instruction
    Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C, Avoid freeze / thaw cycle.
    Host
    Rabbit
    Applications
    WB Image

    Western blot – LC3B antibody (AT0763) at 1/1000 dilution
    Analysis of LC3B in treated U87-MG (human glioblastoma astrocytoma) lysates.
     
    Recommended secondary antibody: Goat anti rabbit IgG-HRP (Engibody, Catalog number:AT0097)
    Recommended ECL: ECL Pico-Detect™ Western Chemiluminescent HRP Substrate (Engibody, Catalog number: IF6747)

    ICC/IF Image

    Genetic Strategies Validation. Knockout Validated: LC3B Antibody - LC3 was detected in immersion fixed Cloroquine treated Hela cells (left) but was not detected in LC3 knockout Hela cells (right) using anti- LC3 rabbit polyclonal antibody at 0.3 ug/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm.

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