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LC3 Antibody (APG8A) (N-term)

Purified Rabbit Polyclonal Antibody (Pab)

     
  • IF - LC3 Antibody (APG8A) (N-term) AP1801A
    Fluorescent image of U251 cells stained with AP1801a 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 AP1801a 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.
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  • WB - LC3 Antibody (APG8A) (N-term) AP1801A
    Western blot analysis of anti-LC3 (APG8a) Pab (Cat. #AP1801a) 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).
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  • IHC-P - LC3 Antibody (APG8A) (N-term) AP1801A
    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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  • SPECIFICATION
  • CITATIONS: 28
  • PROTOCOLS
  • BACKGROUND
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Product Information
Application
  • Applications Legend:
  • WB=Western Blot
  • IHC=Immunohistochemistry
  • IHC-P=Immunohistochemistry (Paraffin-embedded Sections)
  • IHC-F=Immunohistochemistry (Frozen Sections)
  • IF=Immunofluorescence
  • FC=Flow Cytopmetry
  • IC=Immunochemistry
  • ICC=Immunocytochemistry
  • E=ELISA
  • IP=Immunoprecipitation
  • DB=Dot Blot
  • CHIP=Chromatin Immunoprecipitation
  • FA=Fluorescence Assay
  • IEM=Immuno electron microscopy
  • EIA=Enzyme Immunoassay
IHC-P, IF, WB, E
Primary Accession Q9H492
Reactivity Human, Mouse, Rat
Host Rabbit
Clonality Polyclonal
Isotype Rabbit IgG
Calculated MW 14272 Da
Antigen Region 1-30 aa
Additional Information
Gene ID 84557
Other Names 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
Target/Specificity 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).
Dilution IF~~1:200
WB~~1000
IHC-P~~1:50~100
Format 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.
StorageMaintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
PrecautionsLC3 Antibody (APG8A) (N-term) is for research use only and not for use in diagnostic or therapeutic procedures.
Protein Information
Name MAP1LC3A
Function Ubiquitin-like modifier involved in formation of autophagosomal vacuoles (autophagosomes) (PubMed:20713600, PubMed:24290141). While LC3s are involved in elongation of the phagophore membrane, the GABARAP/GATE-16 subfamily is essential for a later stage in autophagosome maturation (PubMed:20713600). Through its interaction with the reticulophagy receptor TEX264, participates in the remodeling of subdomains of the endoplasmic reticulum into autophagosomes upon nutrient stress, which then fuse with lysosomes for endoplasmic reticulum turnover (PubMed:31006537, PubMed:31006538).
Cellular Location Cytoplasmic vesicle, autophagosome membrane; Lipid-anchor. Endomembrane system; Lipid-anchor. Cytoplasm, cytoskeleton {ECO:0000250|UniProtKB:Q91VR7}. Note=LC3-II binds to the autophagic membranes.
Tissue Location Most abundant in heart, brain, liver, skeletal muscle and testis but absent in thymus and peripheral blood leukocytes
Research Areas
Citations ( 0 )

Background

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.

References

References for protein:
1.Baehrecke EH. Nat Rev Mol Cell Biol. 6(6):505-10. (2005)
2.Lum JJ, et al. Nat Rev Mol Cell Biol. 6(6):439-48. (2005)
3.Greenberg JT. Dev Cell. 8(6):799-801. (2005)
4.Levine B. Cell. 120(2):159-62. (2005)
5.Shintani T and Klionsky DJ. Science. 306(5698):990-5. (2004)
6.Tanida I., et al. Int. J. Biochem. Cell Biol. 36:2503-2518(2004)
7.He H., et al. J. Biol. Chem. 278:29278-29287(2003)
8.Tanida I., et al. J. Biol. Chem. 279:36268-36276(2004)
References for U251 cell line:
1. Westermark B.; Pontén J.; Hugosson R. (1973).” Determinants for the establishment of permanent tissue culture lines from human gliomas”. Acta Pathol Microbiol Scand A. 81:791-805. [PMID: 4359449].
2. Pontén, J.,Westermark B. (1978).” Properties of Human Malignant Glioma Cells in Vitro”. Medical Biology 56: 184-193.[PMID: 359950].
3. Geng Y.;Kohli L.; Klocke B.J.; Roth K.A.(2010). “Chloroquine-induced autophagic vacuole accumulation and cell death in glioma cells is p53 independent”. Neuro Oncol. 12(5): 473–481.[ PMID: 20406898].

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Cat# AP1801A
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