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APG5 Antibody

Purified Rabbit Polyclonal Antibody (Pab)

     
  • 1 - APG5 Antibody AP1812C
    Western blot analysis of APG5 (arrow) using rabbit polyclonal APG5 Antibody (Cat. #AP1812c). 293T cell lysates either nontransfected (Lane 1) or transiently transfected (Lane 2) with the APG5 gene.
  • 1 - APG5 Antibody AP1812C
    Western blot analysis of lysate from K562 cell line, using APG5 Antibody(Cat. #AP1812c). AP1812c was diluted at 1:1000. A goat anti-rabbit IgG H&L(HRP) at 1:10000 dilution was used as the secondary antibody. Lysate at 20ug.
  • 3 - APG5 Antibody AP1812C
    Fluorescent image of U251 cells stained with APG5 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 AP1812c APG5 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.
  • 1 - APG5 Antibody AP1812C
    APG5 Antibody (Cat.#AP1812c) western blot analysis in uterus tumor cell line lysates (35ug/lane).This demonstrates the APG5 antibody detected the APG5 protein (arrow).
  • 14 - APG5 Antibody AP1812C
    APG5 Antibody (Cat. #AP1812cb)immunohistochemistry analysis in formalin fixed and paraffin embedded human stomach tissue followed by peroxidase conjugation of the secondary antibody and DAB staining.This data demonstrates the use of APG5 Antibody for immunohistochemistry. Clinical relevance has not been evaluated.
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Product Information
Application
  • Applications Legend:
  • E=ELISA
  • WB=Western Blotting
  • IHC=Immunohistochemistry
  • IHC-P=Immunohistochemistry (Paraffin)
  • IP=Immunoprecipitation
  • IF=Immunofluorescence
  • IC=Immunochemistry
  • ICC=Immunocytochemistry
  • FC=Flow Cytometry
  • DB=Dot Blot
IF, WB, IHC-P, E
Primary Accession Q9H1Y0
Reactivity Human
Host Rabbit
Clonality Polyclonal
Isotype Rabbit IgG
Calculated MW 32447 Da
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 selected from the full length of human ATG5.
Dilution WB~~1:1000
IF~~1:200
IHC-P~~1:100~500
Format Purified monoclonal antibody supplied in PBS with 0.09% (W/V) sodium azide. This antibody is purified through a protein A column, 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.
PrecautionsAPG5 Antibody is for research use only and not for use in diagnostic or therapeutic procedures.
Protein Information
Name ATG5
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.
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

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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