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ATG5 Antibody (N-term)

Purified Rabbit Polyclonal Antibody (Pab)

     
  • IF - ATG5 Antibody (N-term) AP1812a
    Fluorescent image of U251 cells stained with ATG5 (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 AP1812a ATG5 (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). ATG5 immunoreactivity is localized to autophagic vacuoles in the cytoplasm of U251 cells.
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  • WB - ATG5 Antibody (N-term) AP1812a
    All lanes : Anti-hAPG5L-D3 at 1:1000 dilution Lane 1: SH-SY5Y whole cell lysate Lane 2: Hela 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 : 32 kDa Blocking/Dilution buffer: 5% NFDM/TBST.
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  • IHC-P-Leica - ATG5 Antibody (N-term) AP1812a
    Immunohistochemical analysis of paraffin-embedded human liver tissue using AP1812a performed on the Leica® BOND RXm. Tissue was fixed with formaldehyde at room temperature, antigen retrieval was by heat mediation with a EDTA buffer (pH9. 0). Samples were incubated with primary antibody(1:500) for 1 hours at room temperature. A undiluted biotinylated CRF Anti-Polyvalent HRP Polymer antibody was used as the secondary antibody.
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  • WB - ATG5 Antibody (N-term) AP1812a
    The anti-APG5L Pab (Cat. #AP1812a) is used in Western blot to detect APG5L in mouse brain tissue lysate.
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  • SPECIFICATION
  • CITATIONS: 28
  • PROTOCOLS
  • BACKGROUND
  • detail
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
WB, IF, IHC-P-Leica, E
Primary Accession Q9H1Y0
Other Accession Q3MQ06, Q3MQ04, Q99J83, Q3MQ24, Q6DEM4
Reactivity Human, Mouse
Predicted Bovine, Pig, Rat
Host Rabbit
Clonality Polyclonal
Isotype Rabbit IgG
Calculated MW 32447 Da
Antigen Region 1-30 aa
Additional Information
Gene ID 9474
Other Names Autophagy protein 5, APG5-like, Apoptosis-specific protein, ATG5, APG5L, ASP
Target/Specificity This ATG5 antibody is generated from rabbits immunized with a KLH conjugated synthetic peptide between 1-30 amino acids from the N-terminal region of human ATG5.
Dilution IF~~1:200
WB~~1:1000
IHC-P-Leica~~1:500
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.
PrecautionsATG5 Antibody (N-term) is for research use only and not for use in diagnostic or therapeutic procedures.
Protein Information
Name ATG5 (HGNC:589)
Synonyms APG5L, ASP
Function Involved in autophagic vesicle formation. Conjugation with ATG12, through a ubiquitin-like conjugating system involving ATG7 as an E1-like activating enzyme and ATG10 as an E2-like conjugating enzyme, is essential for its function. The ATG12-ATG5 conjugate acts as an E3- like enzyme which is required for lipidation of ATG8 family proteins and their association to the vesicle membranes. Involved in mitochondrial quality control after oxidative damage, and in subsequent cellular longevity. Plays a critical role in multiple aspects of lymphocyte development and is essential for both B and T lymphocyte survival and proliferation. Required for optimal processing and presentation of antigens for MHC II. Involved in the maintenance of axon morphology and membrane structures, as well as in normal adipocyte differentiation. Promotes primary ciliogenesis through removal of OFD1 from centriolar satellites and degradation of IFT20 via the autophagic pathway. As part of the ATG8 conjugation system with ATG12 and ATG16L1, required for recruitment of LRRK2 to stressed lysosomes and induction of LRRK2 kinase activity in response to lysosomal stress (By similarity).
Cellular Location Cytoplasm. Preautophagosomal structure membrane; Peripheral membrane protein Note=Colocalizes with nonmuscle actin. The conjugate detaches from the membrane immediately before or after autophagosome formation is completed (By similarity). Localizes also to discrete punctae along the ciliary axoneme and to the base of the ciliary axoneme.
Tissue Location Ubiquitous. The mRNA is present at similar levels in viable and apoptotic cells, whereas the protein is dramatically highly expressed in apoptotic cells
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). APG5, required for autophagy, conjugates to ATG12 and associates with an isolation membrane to form a cup-shaped isolation membrane and autophagosome. The conjugate detaches from the membrane immediately before or after autophagosome formation is completed. APG5 may also play an important role in the apoptotic process, possibly within the modified cytoskeleton. Its expression is a relatively late event in the apoptotic process, occurring downstream of caspase activity.

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.Hammond E.M., et al. FEBS Lett. 425:391-395(1998)
7. Strausberg R.L., et al. PNAS 99:16899-16903(2002)
8.Grand R.J.A., et al. Exp. Cell Res. 218:439-451(1995)
9.Mizushima N., et al. J. Biol. Chem. 273:33889-33892(1998)
10.Mizushima N., et al. J. Cell Biol. 152:657-668(2001)
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# AP1812a
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