Human/Mouse ATG7 Antibody

(2 citations)   
  • Species Reactivity
    Human, Mouse
  • Specificity
    Detects human ATG7 in direct ELISAs and Western blots. In direct ELISAs and Western blots, no cross-reactivity with recombinant human ATG3, 4A, 4B, 5, 10, 12, or 16L1 is observed.
  • Source
    Monoclonal Mouse IgG1 Clone # 683906
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    E. coli-derived recombinant human ATG7
    Asn387-Gln570
    Accession # O95352
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    2 µg/mL
    See below
  • Simple Western
    20 µg/mL
    See below
  • Immunohistochemistry
    8-25 µg/mL
    See below
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Immunocytochemistry
    8-25 µg/mL
    See below
  • Intracellular Staining by Flow Cytometry
    0.25 µg/106 cells
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Human ATG7 by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line and HepG2 human hepatocellular carcinoma cell line. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human ATG7 Monoclonal Antibody (Catalog # MAB6608) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for ATG7 at approximately 75 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunocytochemistry
ATG7 in RAW 264.7 Mouse Cell Line. ATG7 was detected in immersion fixed RAW 264.7 mouse monocyte/macrophage cell line using Mouse Anti-Human ATG7 Monoclonal Antibody (Catalog # MAB6608) at 25 µg/mL for 3 hours at room temperature. Cells were stained using the Northern­Lights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to autophagosomes. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Immunohistochemistry
ATG7 in Human Brain. ATG7 was detected in formalin fixed paraffin-embedded sections of human brain (cortex) using Mouse Anti-Human ATG7 Monoclonal Antibody (Catalog # MAB6608) at 15 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counter­stained with hematoxylin (blue). Specific staining was localized to neuronal cell bodies and processes. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Intracellular Staining by Flow Cytometry
Detection of ATG7 in HeLa Human Cell Line by Flow Cytometry. HeLa human cervical epithelial carcinoma cell line was stained with Mouse Anti-Human/Mouse ATG7 Monoclonal Antibody
(Catalog # MAB6608, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.
Detection of Human ATG7 by Simple WesternTM. Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line and HepG2 human hepatocellular carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for ATG7 at approximately 66 kDa (as indicated) using 20 µg/mL of Mouse Anti-Human/Mouse ATG7 Monoclonal Antibody (Catalog # MAB6608). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Preparation and Storage
  • Reconstitution
    Sterile PBS to a final concentration of 0.5 mg/mL.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: ATG7

ATG7, also known as APG7, is a 78 kDa cytosolic ubiquitin-E1-like enzyme that plays a key role in the autophagic pathway of intracellular bulk degradation. It is required for the conjugation of ATG5 to ATG12, the lipidation of ATG8, and subsequent autophagosome formation. ATG7 is required for mitochondrial removal during erythropoiesis and for the maintenance of axonal homeostasis. Alternate splicing of human ATG7 generates an isoform that lacks 31 aa at the C-terminus, a region that is required for ATG8 lipidation. Human ATG7 shares 93% aa sequence identitiy with mouse and rat ATG7.

  • Long Name:
    ATG7 Autophagy Related 7 Homolog
  • Entrez Gene IDs:
    74244 (Mouse); 312647 (Rat); 10533 (Yeast); 10533 (Yeast)
  • Alternate Names:
    APG7 autophagy 7-like (S. cerevisiae); APG7; APG7L; APG7-LIKE; ATG12-activating enzyme E1 ATG7; ATG7 autophagy related 7 homolog (S. cerevisiae); ATG7; Autophagy-related protein 7; DKFZp434N0735; GSA7; hAGP7; ubiquitin activating enzyme E1-like protein; Ubiquitin-activating enzyme E1-like protein; ubiquitin-like modifier-activating enzyme ATG7
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

2 Citations: Showing 1 - 2
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Sample Type
  1. Histone deacetylase 3 is required for iNKT cell development
    Authors: P Thapa, S Romero Aro, JY Chung, DB Sant'Angel, VS Shapiro
    Sci Rep, 2017;7(1):5784.
    Species: Mouse
    Sample Type: Whole Cells
    Application: Flow
  2. Autophagy fosters myofibroblast differentiation through MTORC2 activation and downstream upregulation of CTGF.
    Authors: Bernard M, Dieude M, Yang B, Hamelin K, Underwood K, Hebert M
    Autophagy, 2014;10(12):2193-207.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
Cell and Tissue Staining Kits
Description Application Cat# Citations Images  

Anti-Mouse HRP-DAB Cell & Tissue Staining Kit

CTS002 14
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Immunohistochemistry Reagents
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Antigen Retrieval Reagent-Universal

CTS015 10
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Antigen Retrieval Reagent-Basic

CTS013 4
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Mounting Medium

CTS011 4
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Antigen Retrieval Reagent Sampler (50 mL each)

CTS016
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Antigen Retrieval Reagent-Acidic

CTS014
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NorthernLights Guard Mounting Media

NL996
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VisUCyte Antigen Retrieval Reagent-Universal

VCTS023
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VisUCyte Antigen Retrieval Reagent-Basic

VCTS021
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VisUCyte Antigen Retrieval Reagent-Acidic

VCTS022
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Isotype Controls
Description Application Cat# Citations Images  

Mouse IgG1 Isotype Control

Ctrl MAB002 138  
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Mouse IgG1 Isotype Control

Ctrl MAB002R 1
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Secondary Antibodies
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Mouse IgG HRP-conjugated Antibody

WB HAF007 26  
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Mouse F(ab)2 IgG (H+L) PE-conjugated Antibody

Flow F0102B 10  
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Donkey Anti-Mouse IgG NorthernLights™ NL557-conjugate Antibody

Flow, IHC, ICC NL007 9
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Donkey Anti-Mouse IgG Biotinylated Antibody

WB BAF018 1
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Mouse IgG HRP-conjugated Antibody

WB HAF018 9  
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Donkey Anti-Mouse IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL008 8
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Mouse F(ab)2 IgG (H+L) APC-conjugated Antibody

Flow F0101B 2  
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Donkey Anti-Mouse IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL009 5
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Mouse F(ab)2 IgG (H+L) Fluorescein-conjugated Antibody

Flow F0103B 5  
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Goat Anti-Mouse IgG Biotinylated Antibody

WB BAF007 4
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Mouse IgG Antibody

WB AF007 4
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Mouse/Rabbit IgG VisUCyte HRP Polymer Antibody

IHC VC002  
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Mouse IgG VisUCyte HRP Polymer Antibody

IHC VC001  
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Goat Anti-Mouse IgG Fc Affinity Purified Polyclonal Ab

G-202-C 2
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Mouse F(ab)2 IgG (H+L) PerCP-conjugated Antibody

Flow F0114  
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