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ATG5 Antibody (N-term)

Purified Rabbit Polyclonal Antibody (Pab)

     
  • 1 - 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.
  • 3 - 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.
  • 1 - ATG5 Antibody (N-term) AP1812a
    Western blot analysis of APG5L (arrow) using rabbit polyclonal APG5L Antibody (D3) (Cat. #AP1812a). 293 cell lysates (2 ug/lane) either nontransfected (Lane 1) or transiently transfected (Lane 2) with the APG5L gene.
  • 14 - ATG5 Antibody (N-term) AP1812a
    Formalin-fixed and paraffin-embedded human breast carcinoma tissue reacted with Autophagy APG5L antibody (N-term) (Cat.#AP1812a), 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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  • 文献引用 : 25
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Product info
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
WB, IF, IHC-P, E
Primary Accession Q9H1Y0
Other Accession Q3MQ06, Q3MQ04, Q99J83, Q3MQ24
Reactivity Human
Predicted Mouse, Rat, Pig, Bovine
Host Rabbit
Clonality Polyclonal
Isotype Rabbit Ig
Additional info
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 WB~~1:1000
IF~~1:200
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.
PrecautionsATG5 Antibody (N-term) 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. The ATG12- ATG5 conjugate also negatively regulates the innate antiviral immune response by blocking the type I IFN production pathway through direct association with RARRES3 and MAVS. Also plays a role in translation or delivery of incoming viral RNA to the translation apparatus. 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
ATG5 Promotes Death Signaling in Response to the Cyclic Depsipeptides Coibamide A and Apratoxin A.
Author : Wan X1,Serrill JD2,Humphreys IR3,Tan M4,McPhail KL5,Ganley IG6,Ishmael JE7.
Mar Drugs. 2018 Mar 1;16(3). pii: E77. doi: 10.3390/md16030077.
29494533
Atg5flox-Derived Autophagy-Deficient Model of Pompe Disease: Does It Tell the Whole Story?
Author : Lim JA1,2,Zare H1,Puertollano R2,Raben N1.
Mol Ther Methods Clin Dev. 2017 Sep 22;7:11-14. doi: 10.1016/j.omtm.2017.08.002. eCollection 2017 Dec 15.
29057281
The Detection Techniques for Autophagy-Associated Cell Death-Related Genes and Proteins: Gene Expression Assay and Immunohistochemistry.
Author : Ozturk M1,Ozsoylemez OD2,Dagistanli FK3.
Methods Mol Biol. 2017 Sep 10. doi: 10.1007/7651_2017_67. [Epub ahead of print]
28889353
Effects of Combined Lysosomal and Mitochondrial Photodamage in a Non Small-Cell Lung Cancer Cell Line: the Role of Paraptosis.
Author : Kessel D1,Reiners JJ Jr1,2.
Photochem Photobiol. 2017 Jul 11. doi: 10.1111/php.12805. [Epub ahead of print]
28696570
Studying Autophagy in Zebrafish.
Author : Mathai BJ1,Meijer AH2,Simonsen A3.
Cells. 2017 Jul 9;6(3). pii: E21. doi: 10.3390/cells6030021.
28698482
Promotion of Pro-Apoptotic Signals by Lysosomal Photodamage: Mechanistic Aspects and Influence of Autophagy.
Author : Kessel D1, Evans CL2.
Photochem Photobiol. 2016 Apr 20. doi: 10.1111/php.12592. [Epub ahead of print]
27096545
FGFR3/Fibroblast Growth Factor Receptor 3 Inhibits Autophagy through Decreasing the ATG12-ATG5 Conjugate, Leading to the Delay of Cartilage Development in Achondroplasia.
Author : Wang X1,2,3, Qi H1,2,3, Wang Q1,2,3, Zhu Y1,2,3, Wang X1,2,3, Jin M1,2, Autophagy. 2015 Oct 22:0. [Epub ahead of print]
26491898
Autophagy in spinal motor neurons of conditional ADAR2-knockout mice: an implication for a role of calcium in increased autophagy flux in ALS.
Author : Shoichi S1, Yamashita T2, Shin K3.
Neurosci Lett. 2015 May 14. pii: S0304-3940(15)00382-1. doi: 10.1016/j.neulet.2015.05.025. [Epub ahead of print]
25980994
Long-term artificial selection reveals a role of TCTP in autophagy in mammalian cells.
Author : Chen K1, Huang C1, Yuan J1, Cheng H2, Zhou R2.
Mol Biol Evol. 2014 Jun 2. pii: msu181. [Epub ahead of print]
24890374
Coxsackievirus B3 induces crosstalk between autophagy and apoptosis to benefit its release after replicating in autophagosomes through a mechanism involving caspase cleavage of autophagy-related proteins.
Author : Xin L1, Xiao Z1, Ma X1, He F1, Yao H1, Liu Z2.
Infect Genet Evol. 2014 May 15;26C:95-102. doi: 10.1016/j.meegid.2014.05.005. [Epub ahead of print]
24836289
Control of photoreceptor autophagy after retinal detachment: the switch from survival to death.
Author : Chinskey ND1, Zheng QD, Zacks DN.
Invest Ophthalmol Vis Sci. 2014 Feb 4;55(2):688-95. doi: 10.1167/iovs.13-12951.
24408986
Autophagosomes contribute to intracellular lipid distribution in enterocytes.
Author : Khaldoun SA1, Emond-Boisjoly MA, Chateau D, Carrière V, Lacasa M, Rousset M, Demignot S, Morel E.
Mol Biol Cell. 2014 Jan;25(1):118-32. doi: 10.1091/mbc.E13-06-0324. Epub 2013 Oct 30.
24173715
Autophagy Induced by Tumor Necrosis Factor α Mediates Intrinsic Apoptosis in Trophoblastic Cells.
Author : Cha HH, Hwang JR, Kim HY, Choi SJ, Oh SY, Roh CR.
Reprod Sci. 2013 Nov 6. [Epub ahead of print]
24198074
Down-regulation of autophagy-related protein 5 (ATG5) contributes to the pathogenesis of early-stage cutaneous melanoma.
Author : Liu H, He Z, von Rütte T, Yousefi S, Hunger RE, Simon HU.
Sci Transl Med. 2013 Sep 11;5(202):202ra123. doi: 10.1126/scitranslmed.3005864.
24027027
Phosphorylation of Atg5 by the Gadd45β-MEKK4-p38 pathway inhibits autophagy.
Author : Keil E, Höcker R, Schuster M, Essmann F, Ueffing N, Hoffman B, Liebermann DA, Pfeffer K, Schulze-Osthoff K, Schmitz I.
Cell Death Differ. 2013 Feb;20(2):321-32. doi: 10.1038/cdd.2012.129. Epub 2012 Oct 12.
23059785
Cell loss and autophagy in the extra-adrenal chromaffin organ of Zuckerkandl are regulated by glucocorticoid signalling.
Author : Schober A, Parlato R, Huber K, Kinscherf R, Hartleben B, Huber TB, Schütz G, Unsicker K.
J Neuroendocrinol. 2013 Jan;25(1):34-47. doi: 10.1111/j.1365-2826.2012.02367.x.
23078542
Arsenic trioxide enhances the radiation sensitivity of androgen-dependent and -independent human prostate cancer cells.
Author : Chiu HW, Chen YA, Ho SY, Wang YJ.
PLoS One. 2012;7(2):e31579. doi: 10.1371/journal.pone.0031579. Epub 2012 Feb 20.
22363680
Autophagy induced by deficiency of sphingosine-1-phosphate phosphohydrolase 1 is switched to apoptosis by calpain-mediated autophagy-related gene 5 (Atg5) cleavage.
Author : Lépine S, Allegood JC, Edmonds Y, Milstien S, Spiegel S.
J Biol Chem. 2011 Dec 30;286(52):44380-90. doi: 10.1074/jbc.M111.257519. Epub 2011 Nov 3.
22052905
Expression pattern and functions of autophagy-related gene atg5 in zebrafish organogenesis.
Author : Hu Z, Zhang J, Zhang Q.
Autophagy. 2011 Dec;7(12):1514-27.
22082871
Arsenic trioxide induces autophagy and apoptosis in human glioma cells in vitro and in vivo through downregulation of survivin.
Author : Chiu HW, Ho YS, Wang YJ.
J Mol Med (Berl). 2011 Sep;89(9):927-41. doi: 10.1007/s00109-011-0763-1. Epub 2011 May 19.
21594580
A rapid method to improve protein detection by indirect ELISA.
Author : Hnasko R, Lin A, McGarvey JA, Stanker LH.
Biochem Biophys Res Commun. 2011 Jul 15;410(4):726-31. doi: 10.1016/j.bbrc.2011.06.005. Epub 2011 Jun 7.
21679691
p62, Ref(2)P and ubiquitinated proteins are conserved markers of neuronal aging, aggregate formation and progressive autophagic defects.
Author : Bartlett BJ, Isakson P, Lewerenz J, Sanchez H, Kotzebue RW, Cumming RC, Harris GL, Nezis IP, Schubert DR, Simonsen A, Finley KD.
Autophagy. 2011 Jun;7(6):572-83. Epub 2011 Jun 1.
21325881
Sphingosine-1-phosphate phosphohydrolase-1 regulates ER stress-induced autophagy.
Author : Lépine S, Allegood JC, Park M, Dent P, Milstien S, Spiegel S.
Cell Death Differ. 2011 Feb;18(2):350-61. doi: 10.1038/cdd.2010.104. Epub 2010 Aug 27.
20798685
Control of basal autophagy by calpain1 mediated cleavage of ATG5.
Author : Xia HG, Zhang L, Chen G, Zhang T, Liu J, Jin M, Ma X, Ma D, Yuan J.
Autophagy. 2010 Jan;6(1):61-6. Epub 2010 Jan 13.
19901552
An increase in intracellular Ca2+ is required for the activation of mitochondrial calpain to release AIF during cell death.
Author : Norberg E, Gogvadze V, Ott M, Horn M, Uhlén P, Orrenius S, Zhivotovsky B.
Cell Death Differ. 2008 Dec;15(12):1857-64. doi: 10.1038/cdd.2008.123. Epub 2008 Sep 19.
18806756

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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