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ATG16L Antibody

Purified Rabbit Polyclonal Antibody (Pab)

     
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  • IF - ATG16L Antibody AP1817b
    Immunofluorescent analysis of U251 cells, using ATG16L Antibody(Cat. #AP1817b). U251 cells(right) were treated with Chloroquine (50 μM,16h). AP1817b was diluted at 1:25 dilution. Alexa Fluor 488-conjugated goat anti-rabbit lgG at 1:400 dilution was used as the secondary antibody (green).DAPI was used to stain the cell nuclear (blue).
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  • WB - ATG16L Antibody AP1817b
    Cos7, HEK293, MEF, and Hela cells, left to right, respectively. Data courtesy of Drs. Jiefei Geng and Dan Klionsky, University of Michigan.
    detail
  • WB - ATG16L Antibody AP1817b
    Anti-ATG16L Antibody at 1:1000 dilution + Jurkat whole cell lysate Lysates/proteins at 20 µg per lane. Secondary Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/10000 dilution. Predicted band size : 68 kDa Blocking/Dilution buffer: 5% NFDM/TBST.
    detail
  • WB - ATG16L Antibody AP1817b
    APG16L Antibody (Cat. #AP1817b) western blot analysis in NCI-H460 cell line lysates (35ug/lane).This demonstrates the APG16L antibody detected the APG16L protein (arrow).
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  • WB - ATG16L Antibody AP1817b
    Western blot of APG16L (L92) Pab (Cat. #AP1817b) in mouse brain tissue lysate.
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  • IHC-P - ATG16L Antibody AP1817b
    Formalin-fixed and paraffin-embedded human colon carcinoma tissue reacted with Autophagy APG16L antibody (L176), 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.
    detail
  • SPECIFICATION
  • CITATIONS: 12
  • 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=Immunoelectronmicroscopy
  • EIA=Enzyme Immunoassay
IF, WB, IHC-P, E
Primary Accession Q676U5
Reactivity Human, Mouse
Host Rabbit
Clonality Polyclonal
Isotype Rabbit IgG
Calculated MW 68265 Da
Antigen Region 161-190 aa
Additional Information
Gene ID 55054
Other Names Autophagy-related protein 16-1, APG16-like 1, ATG16L1, APG16L
Target/Specificity This ATG16L antibody is generated from rabbits immunized with a KLH conjugated synthetic peptide between 161-190 amino acids from human ATG16L.
Dilution IF~~1:25
WB~~1:1000
IHC-P~~1:10~50
Format Purified polyclonal antibody supplied in PBS with 0.09% (W/V) sodium azide. This antibody is purified through a protein A column, followed by peptide affinity purification.
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.
PrecautionsATG16L Antibody is for research use only and not for use in diagnostic or therapeutic procedures.
Protein Information
Name ATG16L1 {ECO:0000303|PubMed:17200669, ECO:0000312|HGNC:HGNC:21498}
Function Plays an essential role in both canonical and non-canonical autophagy: interacts with ATG12-ATG5 to mediate the lipidation to ATG8 family proteins (MAP1LC3A, MAP1LC3B, MAP1LC3C, GABARAPL1, GABARAPL2 and GABARAP) (PubMed:23376921, PubMed:23392225, PubMed:29317426, PubMed:30778222, PubMed:33909989, PubMed:24553140, PubMed:24954904, PubMed:27273576). Acts as a molecular hub, coordinating autophagy pathways via distinct domains that support either canonical or non- canonical signaling (PubMed:29317426, PubMed:30778222). During canonical autophagy, interacts with ATG12-ATG5 to mediate the conjugation of phosphatidylethanolamine (PE) to ATG8 proteins, to produce a membrane-bound activated form of ATG8 (PubMed:23376921, PubMed:23392225, PubMed:24553140, PubMed:24954904, PubMed:27273576). Thereby, controls the elongation of the nascent autophagosomal membrane (PubMed:23376921, PubMed:23392225, PubMed:24553140, PubMed:24954904, PubMed:27273576). Also involved in non-canonical autophagy, a parallel pathway involving conjugation of ATG8 proteins to single membranes at endolysosomal compartments, probably by catalyzing conjugation of phosphatidylserine (PS) to ATG8 (PubMed:33909989). Non-canonical autophagy plays a key role in epithelial cells to limit lethal infection by influenza A (IAV) virus (By similarity). Regulates mitochondrial antiviral signaling (MAVS)-dependent type I interferon (IFN-I) production (PubMed:22749352, PubMed:25645662). Negatively regulates NOD1- and NOD2-driven inflammatory cytokine response (PubMed:24238340). Instead, promotes an autophagy-dependent antibacterial pathway together with NOD1 or NOD2 (PubMed:20637199). Plays a role in regulating morphology and function of Paneth cell (PubMed:18849966).
Cellular Location Cytoplasm. Preautophagosomal structure membrane; Peripheral membrane protein. Endosome membrane; Peripheral membrane protein. Lysosome membrane; Peripheral membrane protein. Note=Recruited to omegasomes membranes by WIPI2 (By similarity). Omegasomes are endoplasmic reticulum connected strutures at the origin of preautophagosomal structures (By similarity) Localized to preautophagosomal structure (PAS) where it is involved in the membrane targeting of ATG5 (By similarity). Localizes also to discrete punctae along the ciliary axoneme (By similarity). Upon activation of non-canonical autophagy, recruited to single-membrane endolysosomal compartments (PubMed:29317426) {ECO:0000250|UniProtKB:Q8C0J2, ECO:0000269|PubMed:29317426}
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). The APG12-APG5-APG16L complex is esential for the elongation of autophagic isolation membranes. This complex initially associates in uniform distribution with small vesicle membranes. During membrane elongation, the complex partitions, with a great concentration building on the outer side of the isolation membrane. Upon completion of the formation of the autophagosome, the APG12-APG5-APG16L dissociates from the membrane.

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)
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# AP1817b
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