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LC3 Antibody (APG8A) (N-term)

APR06975G Product Datasheet Protocol

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APR06975G		100 μl / $545

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Leading Biology Inc.

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LC3 Antibody (APG8A) (N-term)

Brand	Leading Biology	Catalog Number	APR06975G
Product Type	Polyclonal Antibodies	Field of Research
Product Overview	We constantly strive to ensure we provide our customers with the best antibodies. As a result of this work we offer this antibody in purified format. We are in the process of updating our datasheets. If you have any questions regarding this update, please feel free to contact our technical support team. This product is a high quality LC3 antibody (APG8A) (N-term).
Molecular Weight	14272 Da
Cellular Localization	Antigen Cellular Localization: Cytoplasm, cytoskeleton. Endomembrane system; Lipid-anchor. Cytoplasmic vesicle, autophagosome membrane; Lipid-anchor. Note=LC3-II binds to the autophagic membranes
Host	Rabbit
Species Reactivity	Human, Mouse, Rat
Immunogen	1-30 aa
Target	This LC3 antibody is generated from rabbits immunized with a KLH conjugated synthetic peptide between 1~30 amino acids from the N-term of human LC3 (APG8a).
Isotype	Rabbit Ig
GeneID	84557
UniProt ID	Q9H492
Function	Ubiquitin-like modifier involved in formation of autophagosomal vacuoles (autophagosomes). Whereas LC3s are involved in elongation of the phagophore membrane, the GABARAP/GATE-16 subfamily is essential for a later stage in autophagosome maturation.
Summary	Macroautophagy is the major inducible pathway for the general turnover of cytoplasmic constituents in eukaryotic cells, it is also responsible for the degradation of active cytoplasmic enzymes and organelles during nutrient starvation. Macroautophagy involves the formation of double-membrane bound autophagosomes which enclose the cytoplasmic constituent targeted for degradation in a membrane bound structure, which then fuse with the lysosome (or vacuole) releasing a single-membrane bound autophagic bodies which are then degraded within the lysosome (or vacuole). MAP1A and MAP1B are microtubule-associated proteins which mediate the physical interactions between microtubules and components of the cytoskeleton. These proteins are involved in formation of autophagosomal vacuoles (autophagosomes). MAP1A and MAP1B each consist of a heavy chain subunit and multiple light chain subunits. MAP1LC3a is one of the light chain subunits and can associate with either MAP1A or MAP1B. The precursor molecule is cleaved by APG4B/ATG4B to form the cytosolic form, LC3-I. This is activated by APG7L/ATG7, transferred to ATG3 and conjugated to phospholipid to form the membrane-bound form, LC3-II.
Form	Purified polyclonal antibody supplied in PBS with 0.09% (W/V) sodium azide. This antibody is prepared by Saturated Ammonium Sulfate (SAS) precipitation followed by dialysis against PBS.
Storage & Stability	Store at +4°C short term. For long-term storage, aliquot and store at -20°C or below. Stable for 12 months at -20°C. Avoid repeated freeze-thaw cycles.
Applications	WB, IHC-P, IF, E
Dilution	WB~~1000 IF~~1:200 IHC-P~~1:50~100
Synonyms	Microtubule-associated proteins 1A/1B light chain 3A, Autophagy-related protein LC3 A, Autophagy-related ubiquitin-like modifier LC3 A, MAP1 light chain 3-like protein 1, MAP1A/MAP1B light chain 3 A, MAP1A/MAP1B LC3 A, Microtubule-associated protein 1 light chain 3 alpha, MAP1LC3A
Images	Western blot analysis of anti-LC3 (APG8a) Pab (Cat. APR06975G) in rat brain lysate. Both lipidated (arrow, II) and non-lipidated APG8a (arrow, I) were detected in membrane fraction (P) but only non-lipidated LC3 was detected in soluble fraction (S). Fluorescent image of U251 cells stained with APR06975G LC3 (APG8A) (N-term) antibody. U251 cells were treated with Chloroquine (50 μM,16h), then fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.2%, 30 min). Cells were then incubated with APR06975G LC3 (APG8A) (N-term) primary antibody (1:200, 2 h at room temperature). For secondary antibody, Alexa Fluor? 488 conjugated donkey anti-rabbit antibody (green) was used (1:1000, 1h). Nuclei were counterstained with Hoechst 33342 (blue) (10 μg/ml, 5 min). LC3 immunoreactivity is localized to autophagic vacuoles in the cytoplasm of U251 cells. Formalin-fixed and paraffin-embedded human brain tissue reacted with Autophagy LC3 Antibody (APG8a) (N-term), which was peroxidase-conjugated to the secondary antibody, followed by DAB staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated.
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