Rabbit ATG5 Polyclonal Antibody | anti-ATG5 antibody

ATG5 Antibody (N-term)

Cat. Num. AAA9206419

• Gene Names: ATG5; ASP; APG5; APG5L; hAPG5; APG5-LIKE
• Reactivity: Human;; (Predicted Reactivity: Zebrafish, Bovine, Mouse, Pig, Rat, Rabbit )
• Applications: Western Blot, ELISA, Immunohistochemistry, Immunofluorescence
• Purity: Purified Rabbit Polyclonal Antibody (Pab)
• Synonyms: ATG5; Polyclonal Antibody; ATG5 Antibody (N-term); Autophagy protein 5; APG5-like; Apoptosis-specific protein; APG5L; ASP; anti-ATG5 antibody

• Host: Rabbit
• Reactivity: Human;; (Predicted Reactivity: Zebrafish, Bovine, Mouse, Pig, Rat, Rabbit )
• Clonality: Polyclonal
• Isotype: Rabbit Ig
• 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.
• Purity/Purification: Purified Rabbit Polyclonal Antibody (Pab)
• Form/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.
• Sequence Positions: 1-30
• Sequence Length: 275

• Applicable Applications for anti-ATG5 antibody: Western Blot (WB), ELISA (EIA), Immunohistochemistry-Paraffin (IHC-P), Immunofluorescence (IF)
• Application Notes: WB~~1:1000; IF~~1:200; IHC-P~~1:50~100
• 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 membraneimmediately 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
• Preparation and Storage: Maintain refrigerated at 2-8 degree C for up to 2 weeks. For long term storage store at -20 degree C in small aliquots to prevent freeze-thaw cycles.

Western Blot

(Western blot analysis of lysate from SH-SY5Y cell line, using APG5L Antibody (D3). AP1812a 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.)

Western Blot (WB)

( All lanes : Anti-hAPG5L-D3 at 1:1000 dilutionLane 1: SH-SY5Y whole cell lysate Lane 2: Hela whole cell lysate Lysates/proteins at 20 ug 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.)

Fluorescent

(Fluorescent image of U251 cells stained with ATG5 (N-term) antibody. U251 cells were treated with Chloroquine (50 uM,16h), then fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.2%, 30 min). Cells were then incubated with 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) (10ug/ml, 5 min). ATG5 immunoreactivity islocalized to autophagic vacuoles in the cytoplasm of U251 cells.)

Immunohistochemistry

(Formalin-fixed and paraffin-embedded human breast carcinoma tissue reacted with Autophagy APG5L antibody (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.)

Immunohistochemistry (IHC)

(Immunohistochemical analysis of paraffin-embedded human liver tissue using MBS9206419 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.)

Related Product Information for anti-ATG5 antibody

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.

Product Categories/Family for anti-ATG5 antibody

Crown Antibodies; Cancer; Cell Biology; Developmental Biology; Signal Transduction; Phospho Antibodies

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].

NCBI and Uniprot Product Information

• NCBI GI #: 4757798
• NCBI GeneID: 9474
• NCBI Accession #: NP_004840.1
• NCBI GenBank Nucleotide #: NM_004849.3
• UniProt Accession #: Q9H1Y0; O60875; Q5JVR2; Q68DI4; Q9H2B8; Q9HCZ7
• NCBI Official Full Name: autophagy protein 5 isoform a
• NCBI Official Synonym Full Names: autophagy related 5
• NCBI Official Symbol: ATG5
• NCBI Official Synonym Symbols: ASP; APG5; APG5L; hAPG5; APG5-LIKE
• NCBI Protein Information: autophagy protein 5
• UniProt Protein Name: Autophagy protein 5
• Protein Family: Autophagy protein
• UniProt Gene Name: ATG5
• UniProt Synonym Gene Names: APG5L; ASP
• UniProt Entry Name: ATG5_HUMAN

Uniprot Description

ATG5: Required for autophagy. Conjugates to ATG12 and associates with isolation membrane to form cup-shaped isolation membrane and autophagosome. The conjugate detaches from the membrane immediately before or after autophagosome formation is completed. The ATG5-ATG12 conjugate forms a complex with several units of ATG16. Interacts with TECPR1; the interaction is direct and does not take place when ATG16 is associated with the ATG5- ATG12 conjugate. By apoptotic stimuli. Ubiquitous. The mRNA is present at similar levels in viable and apoptotic cells, whereas the protein is dramatically highly expressed in apoptotic cells. Belongs to the ATG5 family. 2 isoforms of the human protein are produced by alternative splicing.

Protein type: Ubiquitin conjugating system; Apoptosis; Autophagy

Chromosomal Location of Human Ortholog: 6q21

Cellular Component: membrane; cytoplasm; pre-autophagosomal structure membrane; autophagic vacuole; axoneme

Molecular Function: protein binding; APG8 conjugating enzyme activity

Biological Process: response to drug; response to fungus; apoptosis; ventricular cardiac muscle cell development; C-terminal protein lipidation; post-translational protein modification; vasodilation; regulation of release of sequestered calcium ion into cytosol; cellular response to nitrogen starvation; heart contraction; regulation of cytokine secretion during immune response; mitochondrion degradation; innate immune response; autophagy; blood vessel remodeling; negative regulation of protein ubiquitination; negative regulation of interferon type I production; otolith development; autophagic vacuole formation; negative regulation of apoptosis