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ATG5 Antibody (YA615)

(Synonyms: APG 5L; APG5; APG5 autophagy 5 like; APG5 like; APG5-like; Apoptosis specific protein; ASP; ATG 5; ATG5 autophagy related 5 homolog; Autophagy protein 5; hAPG5)
Customer Review

Based on 1 Customer Validation

ATG5 Antibody (YA615) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to ATG5.

For research use only. We do not sell to patients.
  • Host:

    Rabbit

  • Isotype:

    IgG

  • Application:

    WB, ICC/IF, IP

  • Reactivity :

    Human, Mouse, Rat

  • Formulation:

    Supplied in 50 mM Tris-Glycine (pH 7.4), 0.15 M NaCl, 40% Glycerol and 0.05% BSA. Preservative: 0.01% Sodium azide

  • Conjugation:
    Non-conjugated
ALL WB ICC
  • Experimental Validation Results for ATG5 Antibody (YA615)
  • Experimental Validation Results for ATG5 Antibody (YA615)
  • Experimental Validation Results for ATG5 Antibody (YA615)
  • Experimental Validation Results for ATG5 Antibody (YA615)
  • ATG5 Antibody (YA615) Product Image
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Tel: 609-228-6898 E-mail: [email protected] [email protected]

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Related Products:
Top Publications Citing

Use of MCE Products

Applications

Application
WB Info
WB: Western Blot
ICC/IF Info
ICC/IF: Immunocytochemistry/
Immunofluorescence
IP Info
IP: Immunoprecipitation
Dilution Ratio 1:500-1:1000 1:50-1:200 1:20

Product Details

Description

ATG5 Antibody (YA615) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to ATG5.

  • Host Rabbit
  • Clonality Recombinant,Monoclonal
  • Species Reactivity
    Human, Mouse, Rat
  • Observed Molecular Weight
    Observed band size: 55 kDa Info
    Note: Due to possible protein modifications or aggregation, the molecular weight should be confirmed by actual measurement, and the predicted value is for reference only.
  • Calculated Molecular Weight Predicted band size: 32 kDa；
Species Reactivity Database
Immunogen

Synthetic peptide corresponding to Human APG5L/ATG5 aa21-34.

Sensitivity

Endogenous

Purification

affinity purified

Conjugation

Non-conjugated

Modification

Unmodified

Isotype

IgG

RRID

AB_3102601

Product Properties

  • Appearance

    Solution

  • Formulation

    Supplied in 50 mM Tris-Glycine (pH 7.4), 0.15 M NaCl, 40% Glycerol and 0.05% BSA. Preservative: 0.01% Sodium azide

  • Concentration

    Batch-dependent, Please check the COA for the concentration of each lot. Check Lot Concentration

  • Storage & Stability

    Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.

  • Shipping

    Shipping with blue ice.

Verification Images

  • Experimental Validation Results for ATG5 Antibody (YA615)
    Western blot analysis of extracts from Raji(lane 2(20μg) ), Hela(lane 3(20μg) ) ,RAW264.7(lane 4(20μg) ) and NIH3T3(lane 5(20μg)) using APG5L(HY-P80545) Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in TBST for 2 hour at room temperature. The primary antibody (HY-P80545, 1/1000) and Loading control antibody (GAPDH, HY-P80954, 1/3000) was used in 5% BSA in TBST at 4°C overnight. Goat Anti-Rabbit/Mouse IgG-HRP Secondary Antibody (HY-P8001/HY-P8004, 1/10,000) was used for 1 hour at room temperature.
  • Experimental Validation Results for ATG5 Antibody (YA615)
    Western blot analysis was performed on protein extracts (25 μg) from Raji (lane 2), HeLa (lane 3), NIH/3T3 (lane 4), 293T (lane 5), and U87 (lane 6) using ATG5 antibody. Proteins were transferred onto a 0.45 μm PVDF membrane using the Trans-Blot® Turbo™ system for 13 min. The membrane was then blocked with 5% nonfat milk in TBST (HY-K1025) for 1 h at room temperature. The primary antibody (1:1000) and loading control antibody GAPDH Antibody (HRP) (HY-P80954A) (1:5000) were diluted in 5% nonfat milk in TBST and incubated with the membrane overnight at 4°C. After washing, the membrane of primary antibody was incubated with HRP-conjugated goat anti-rabbit/mouse IgG secondary antibody (HY-P8001/HY-P8004) (1:5000) diluted in 5% nonfat milk in TBST for 1 h at room temperature. Protein bands were visualized using an Ultra High Sensitivity ECL detection kit (HY-K1005).
  • Experimental Validation Results for ATG5 Antibody (YA615)
    Immunocytochemistry analysis of MCF-7 cells labeling ATG5 with ATG5 Antibody (HY-P80545) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with quick block buffer for 10 minutes at room temperature. Cells were then incubated with ATG5 Antibody (HY-P80545) at 1/100 dilution in quick block buffer overnight at 4 ℃. AF488-conjugated Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
  • Experimental Validation Results for ATG5 Antibody (YA615)
    Immunocytochemistry analysis of NIH/3T3 cells labeling ATG5 with ATG5 Antibody (HY-P80545) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with quick block buffer for 10 minutes at room temperature. Cells were then incubated with ATG5 Antibody (HY-P80545) at 1/100 dilution in quick block buffer overnight at 4 ℃. AF488-conjugated Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).

Background

  • Function

    ATG5 is 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; May 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. Plays a crucial role in IFN-gamma-induced autophagic cell death by interacting with FADD; (Microbial infection) May act as a proviral factor. In association with ATG12, negatively regulates the innate antiviral immune response by impairing the type I IFN production pathway upon vesicular stomatitis virus (VSV) infection. Required for the translation of incoming hepatitis C virus (HCV) RNA and, thereby, for initiation of HCV replication, but not required once infection is established[1][2].

  • Subcellular Localization

    Cytoplasm; Preautophagosomal structure membrane; Peripheral membrane protein

  • Expression


    Tissue_specificity:It is ubiquitous. The levels of mRNA are similar in living cells and apoptotic cells, while protein expression is significantly elevated in apoptotic cells.

    Induction:By apoptotic stimuli

  • Isoforms & Post-Translational Modification

    Q9H1Y0 has 2 isomers: Q9H1Y0-1: 32447 Da (predicted); Q9H1Y0-2: 22931 Da (predicted).
    Conjugated to ATG12; which is essential for autophagy, but is not required for association with isolation membrane;Acetylated by EP300

  • Subunit

    Forms a conjugate with ATG12 (PubMed:11825910, PubMed:12207896, PubMed:17709747, PubMed:26812546). Part of the minor complex composed of 4 sets of ATG12-ATG5 and ATG16L1 (400 kDa); this complex interacts with ATG3 leading to disruption of ATG7 interaction and promotion of ATG8-like proteins lipidation (PubMed:24191030). Forms an 800-kDa complex composed of ATG12-ATG5 and ATG16L2 (By similarity). The ATG12-ATG5 conjugate interacts with RAB33A; this interaction is bridged by ATG16L1 and promotes ATG12-ATG5-ATG16L1 complex recruitment to phagophores (PubMed:32960676). Interacts with TECPR1; the interaction is direct and does not take place when ATG16L1 is associated with the ATG5-ATG12 conjugate (PubMed:21575909, PubMed:22342342). Interacts with DHX58/RIG-1, IFIH1/MDA5 and MAVS/IPS-1 in monomeric form as well as in ATG12-ATG5 conjugate form. The interaction with MAVS is further enhanced upon vesicular stomatitis virus (VSV) infection (PubMed:17709747). Interacts with ATG3 (PubMed:11825910, PubMed:12207896). Interacts with ATG7 and ATG10 (By similarity). Interacts with FADD (PubMed:15778222). Interacts with Bassoon/BSN; this interaction is important for the regulation of presynaptic autophagy (By similarity). Interacts with ATG16L2 (By similarity)

  • SwissProt ID

    Q9H1Y0

  • Gene ID
    9474 [NCBI]
  • Synonyms

    APG 5L; APG5; APG5 autophagy 5 like; APG5 like; APG5-like; Apoptosis specific protein; ASP; ATG 5; ATG5 autophagy related 5 homolog; Autophagy protein 5; hAPG5

  • Research Field

    Cell Biology

[1]. Jounai N, et al. The Atg5 Atg12 conjugate associates with innate antiviral immune responses. Proc Natl Acad Sci U S A. 2007 Aug 28;104(35):14050-5. [Content Brief]

[2]. Dreux M, et al. The autophagy machinery is required to initiate hepatitis C virus replication. Proc Natl Acad Sci U S A. 2009 Aug 18;106(33):14046-51. [Content Brief]

ATG5 Antibody (YA615) Related Classifications

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100 mg

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Measured by its ability to inhibit anti-CD3-induced proliferation of stimulated CTLL-2 mouse cytotoxic T cells. The ED50 this effect is 0.8023 μg/ml, corresponding to a specific activity is 1246.42 units/mg.

Assay Types
  • 1/16

    MedchemExpress Validation
     Western blot analysis of extracts from Raji(lane 2(20μg) ), Hela(lane 3(20μg) ) ,RAW264.7(lane 4(20μg) ) and NIH3T3(lane 5(20μg)) using APG5L(HY-P80545) Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in TBST for 2 hour at room temperature. The primary antibody (HY-P80545, 1/1000) and Loading control antibody (GAPDH, HY-P80954, 1/3000) was used in 5% BSA in TBST at 4°C overnight. Goat Anti-Rabbit/Mouse IgG-HRP Secondary Antibody (HY-P8001/HY-P8004, 1/10,000) was used for 1 hour at room temperature.

  • 1/16

    MedChemExpress Validation
     Western blot analysis was performed on protein extracts (25 μg) from Raji (lane 2), HeLa (lane 3), NIH/3T3 (lane 4), 293T (lane 5), and U87 (lane 6) using ATG5 antibody. Proteins were transferred onto a 0.45 μm PVDF membrane using the Trans-Blot® Turbo™ system for 13 min. The membrane was then blocked with 5% nonfat milk in TBST (HY-K1025) for 1 h at room temperature. The primary antibody (1:1000) and loading control antibody GAPDH Antibody (HRP) (HY-P80954A) (1:5000) were diluted in 5% nonfat milk in TBST and incubated with the membrane overnight at 4°C. After washing, the membrane of primary antibody was incubated with HRP-conjugated goat anti-rabbit/mouse IgG secondary antibody (HY-P8001/HY-P8004) (1:5000) diluted in 5% nonfat milk in TBST for 1 h at room temperature. Protein bands were visualized using an Ultra High Sensitivity ECL detection kit (HY-K1005).

  • 1/16

    MedChemExpress Validation
     Immunocytochemistry analysis of MCF-7 cells labeling ATG5 with ATG5 Antibody (HY-P80545) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with quick block buffer for 10 minutes at room temperature. Cells were then incubated with ATG5 Antibody (HY-P80545) at 1/100 dilution in quick block buffer overnight at 4 ℃. AF488-conjugated Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).

  • 1/16

    MedChemExpress Validation
     Immunocytochemistry analysis of NIH/3T3 cells labeling ATG5 with ATG5 Antibody (HY-P80545) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with quick block buffer for 10 minutes at room temperature. Cells were then incubated with ATG5 Antibody (HY-P80545) at 1/100 dilution in quick block buffer overnight at 4 ℃. AF488-conjugated Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).

  • ATG5 Antibody (YA615) Product Image
    1/16
  • Experimental Validation Results for ATG5 Antibody (YA615)
  • Experimental Validation Results for ATG5 Antibody (YA615)
  • Experimental Validation Results for ATG5 Antibody (YA615)
  • Experimental Validation Results for ATG5 Antibody (YA615)
  • ATG5 Antibody (YA615) Product Image
  • Experimental Validation Results for ATG5 Antibody (YA615)
    1/1
    MedchemExpress Validation
     Western blot analysis of extracts from Raji(lane 2(20μg) ), Hela(lane 3(20μg) ) ,RAW264.7(lane 4(20μg) ) and NIH3T3(lane 5(20μg)) using APG5L(HY-P80545) Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in TBST for 2 hour at room temperature. The primary antibody (HY-P80545, 1/1000) and Loading control antibody (GAPDH, HY-P80954, 1/3000) was used in 5% BSA in TBST at 4°C overnight. Goat Anti-Rabbit/Mouse IgG-HRP Secondary Antibody (HY-P8001/HY-P8004, 1/10,000) was used for 1 hour at room temperature.
  • Experimental Validation Results for ATG5 Antibody (YA615)
    1/1
    MedChemExpress Validation
     Western blot analysis was performed on protein extracts (25 μg) from Raji (lane 2), HeLa (lane 3), NIH/3T3 (lane 4), 293T (lane 5), and U87 (lane 6) using ATG5 antibody. Proteins were transferred onto a 0.45 μm PVDF membrane using the Trans-Blot® Turbo™ system for 13 min. The membrane was then blocked with 5% nonfat milk in TBST (HY-K1025) for 1 h at room temperature. The primary antibody (1:1000) and loading control antibody GAPDH Antibody (HRP) (HY-P80954A) (1:5000) were diluted in 5% nonfat milk in TBST and incubated with the membrane overnight at 4°C. After washing, the membrane of primary antibody was incubated with HRP-conjugated goat anti-rabbit/mouse IgG secondary antibody (HY-P8001/HY-P8004) (1:5000) diluted in 5% nonfat milk in TBST for 1 h at room temperature. Protein bands were visualized using an Ultra High Sensitivity ECL detection kit (HY-K1005).
  • Experimental Validation Results for ATG5 Antibody (YA615)
    1/1
    MedChemExpress Validation
     Immunocytochemistry analysis of MCF-7 cells labeling ATG5 with ATG5 Antibody (HY-P80545) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with quick block buffer for 10 minutes at room temperature. Cells were then incubated with ATG5 Antibody (HY-P80545) at 1/100 dilution in quick block buffer overnight at 4 ℃. AF488-conjugated Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
  • Experimental Validation Results for ATG5 Antibody (YA615)
    1/1
    MedChemExpress Validation
     Immunocytochemistry analysis of NIH/3T3 cells labeling ATG5 with ATG5 Antibody (HY-P80545) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with quick block buffer for 10 minutes at room temperature. Cells were then incubated with ATG5 Antibody (HY-P80545) at 1/100 dilution in quick block buffer overnight at 4 ℃. AF488-conjugated Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
  • Experimental Validation Results for ATG5 Antibody (YA615)
  • Experimental Validation Results for ATG5 Antibody (YA615)
  • Experimental Validation Results for ATG5 Antibody (YA615)
  • Experimental Validation Results for ATG5 Antibody (YA615)
  • ATG5 Antibody (YA615) Product Image
    MedchemExpress Validation

    Western blot analysis of extracts from Raji(lane 2(20μg) ), Hela(lane 3(20μg) ) ,RAW264.7(lane 4(20μg) ) and NIH3T3(lane 5(20μg)) using APG5L(HY-P80545) Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in TBST for 2 hour at room temperature. The primary antibody (HY-P80545, 1/1000) and Loading control antibody (GAPDH, HY-P80954, 1/3000) was used in 5% BSA in TBST at 4°C overnight. Goat Anti-Rabbit/Mouse IgG-HRP Secondary Antibody (HY-P8001/HY-P8004, 1/10,000) was used for 1 hour at room temperature.

  • ATG5 Antibody (YA615) Product Image
    MedChemExpress Validation

    Western blot analysis was performed on protein extracts (25 μg) from Raji (lane 2), HeLa (lane 3), NIH/3T3 (lane 4), 293T (lane 5), and U87 (lane 6) using ATG5 antibody. Proteins were transferred onto a 0.45 μm PVDF membrane using the Trans-Blot® Turbo™ system for 13 min. The membrane was then blocked with 5% nonfat milk in TBST (HY-K1025) for 1 h at room temperature. The primary antibody (1:1000) and loading control antibody GAPDH Antibody (HRP) (HY-P80954A) (1:5000) were diluted in 5% nonfat milk in TBST and incubated with the membrane overnight at 4°C. After washing, the membrane of primary antibody was incubated with HRP-conjugated goat anti-rabbit/mouse IgG secondary antibody (HY-P8001/HY-P8004) (1:5000) diluted in 5% nonfat milk in TBST for 1 h at room temperature. Protein bands were visualized using an Ultra High Sensitivity ECL detection kit (HY-K1005).

  • ATG5 Antibody (YA615) Product Image
    MedChemExpress Validation

    Immunocytochemistry analysis of MCF-7 cells labeling ATG5 with ATG5 Antibody (HY-P80545) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with quick block buffer for 10 minutes at room temperature. Cells were then incubated with ATG5 Antibody (HY-P80545) at 1/100 dilution in quick block buffer overnight at 4 ℃. AF488-conjugated Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).

  • ATG5 Antibody (YA615) Product Image
    MedChemExpress Validation

    Immunocytochemistry analysis of NIH/3T3 cells labeling ATG5 with ATG5 Antibody (HY-P80545) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with quick block buffer for 10 minutes at room temperature. Cells were then incubated with ATG5 Antibody (HY-P80545) at 1/100 dilution in quick block buffer overnight at 4 ℃. AF488-conjugated Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).

  • ATG5 Antibody (YA615) Product Image WB
  • ATG5 Antibody (YA615) Product Image WB
  • ATG5 Antibody (YA615) Product Image ICC
  • ATG5 Antibody (YA615) Product Image ICC