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Purified anti-ATG7 Antibody

Pricing & Availability
Clone
A17129A (See other available formats)
Regulatory Status
RUO
Other Names
APG7, APG7-like, APG7L, Ubiquitin-activating enzyme E1-like protein, Ubiquitin-Like Modifier-Activating Enzyme ATG7, Autophagy-related protein 7
Isotype
Mouse IgG2b, κ
Ave. Rating
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Product Citations
publications
A. A17129A_PURE_ATG7_WB_101723
Whole tissue or cell extracts (15 µg total protein) were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 1.0 µg/mL of purified anti-ATG7 (clone A17129A) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse IgG (Cat. No. 405306) at a 1:3000 dilution. Direct-Blot™ HRP anti-GAPDH (Cat. No. 607903) was used as a loading control at a 1:50000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Lane M: Molecular weight marker
  • A. A17129A_PURE_ATG7_WB_101723
    Whole tissue or cell extracts (15 µg total protein) were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 1.0 µg/mL of purified anti-ATG7 (clone A17129A) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse IgG (Cat. No. 405306) at a 1:3000 dilution. Direct-Blot™ HRP anti-GAPDH (Cat. No. 607903) was used as a loading control at a 1:50000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Lane M: Molecular weight marker
  • B. A17129A_PURE_ATG7_ICC_101723
    U-251 MG cells were fixed with Fixation Buffer (Cat. No. 420801) for 10 minutes and permeabilized with 0.5% Triton X-100 for 10 minutes, and blocked with 5% FBS for 1 hour at room temperature. Cells were then stained with 10 µg/mL of purified anti-ATG7 (clone A17129A), followed by incubation with Alexa Fluor® 647 Goat anti-mouse IgG (Cat. No. 405322) for 1 hour at room temperature (panel A). Cells were also stained with Flash Phalloidin™ Green 488 (Cat. No. 424201) for 1 hour at room temperature and nuclei were counterstained with DAPI (blue) (Cat. No. 422801) (merge in panel B). The images were captured with a 60X objective. Scale bar: 50 µm
  • C. A17129A_PURE_ATG7_ICC_10-17-23
    Rat primary neurons were fixed with Fixation Buffer (Cat. No. 420801) for 10 minutes and permeabilized with 0.5% Triton X-100 for 10 minutes, and blocked with 5% FBS for 1 hour at room temperature. Cells were then stained with 10 µg/mL of purified anti-ATG7 (clone A17129A), followed by incubation with Alexa Fluor® 647 goat anti-mouse IgG (Cat. No. 405322) for 1 hour at room temperature (panel A). Cells were also stained with Flash Phalloidin™ Green 488 (Cat. No. 424201) for 1 hour at room temperature and nuclei were counterstained with DAPI (blue) (Cat. No. 422801) (merge in panel B). The images were captured with a 60X objective. Scale bar: 50 µm
  • D. A17129A_PURE_ATG7_IHC-P_10-17-23
    IHC staining of purified anti-ATG7 (clone A17129A) on formalin-fixed paraffin-embedded human brain tissue. Following antigen retrieval using Citrate Buffer, 10X (Cat. No. 420902), the tissue was incubated with 5 µg/mL of the primary antibody overnight at 4°C. BioLegend’s Ultra Streptavidin HRP Kit (Multi-Species, DAB, Cat. No. 929501) was used for detection followed by hematoxylin counterstaining, according to the protocol provided. Images were captured with a 40X objective. Scale bar: 50 µm
  • E. A17129A_PURE_ATG7_IHCP_10-17-23
    IHC staining of purified anti-ATG7 (clone A17129A) on formalin-fixed paraffin-embedded mouse brain tissue. Following antigen retrieval using Citrate Buffer, 10X (Cat. No. 420902), the tissue was incubated with 10 µg/mL of the primary antibody overnight at 4°C. BioLegend’s Ultra Streptavidin HRP Kit (Multi-Species, DAB, Cat. No. 929501) was used for detection followed by hematoxylin counterstaining, according to the protocol provided. Images were captured with a 40X objective. Scale bar: 50 µm
  • F. A17129A_PURE_ATG7_IHCP_101723
    IHC staining of purified anti-ATG7 (clone A17129A) on formalin-fixed paraffin-embedded rat brain tissue. Following antigen retrieval using Citrate Buffer, 10X (Cat. No. 420902), the tissue was incubated with 5 µg/mL of the primary antibody overnight at 4°C. BioLegend’s Ultra Streptavidin HRP Kit (Multi-Species, DAB, Cat. No. 929501) was used for detection followed by hematoxylin counterstaining, according to the protocol provided. Images were captured with a 40X objective. Scale bar: 50 µm
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625851 25 µg $170
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625852 100 µg $425
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Description

ATG7 is an E1-like ligase that plays a central role in autophagosome biogenesis by conjugating ATG5 to ATG12 which degrades and recycles damaged organelles, protein aggregates, and invading pathogens. ATG7 interacts with other ATG proteins, such as ATG3, ATG10, and ATG16L1, to form a complex that is required for autophagy, also regulates immunity, cell death, and protein secretion, and independently regulates the cell cycle and apoptosis. It also participates in lipidation of the ATG8 protein, which is essential for autophagosome formation. Additionally, ATG7 has been shown to interact with other proteins outside of the autophagy pathway, such as p53, to regulate apoptosis. Disrupting ATG7-dependent autophagy has been shown to cause electromotility disturbances, outer hair cell loss, and deafness in mice. ATG7 dysfunction has been associated with various human diseases, including cancer, neurodegeneration, and infection. ATG7 dysfunction and disease was found in patients with biallelic ATG7 variants and childhood-onset neuropathology. Furthermore, ATG7 has been shown to induce basal autophagy and rescue autophagic deficiency in CryABR120G cardiomyocytes.

Product Details
Technical data sheet

Product Details

Verified Reactivity
Human, Mouse, Rat
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Recombinant fragment of human ATG7
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

WB - Quality tested
ICC, IHC-P - Verified
Recommended Usage

Each lot of this antibody is quality control tested by western blotting. For western blotting, the suggested use of this reagent is 0.1 - 1.0 µg/mL. For immunocytochemistry, a concentration range of 5.0 - 10.0 μg/mL is recommended. For immunohistochemistry on formalin-fixed paraffin-embedded tissue sections, a concentration range of 5.0 - 10.0 µg/mL is suggested. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

For immunohistochemistry (IHC-P), Citrate Buffer, 10X (Cat. No, 420902) is recommended.

For immunocytochemistry (ICC), either Fixation Buffer (Cat. No. 420801) or paraformaldehyde-based fixation is recommended.

Clone A17129A detects homodimer of ATG7 in western blotting (WB). The homodimer band is demolished by increasing DTT concentration in the lysis buffer.

RRID
AB_3083415 (BioLegend Cat. No. 625851)
AB_3083415 (BioLegend Cat. No. 625852)

Antigen Details

Structure
Homodimer
Distribution

Cytosol and preautophagosomal structure

Function
Conjugate ATG12 to ATG5, lipidation of ATG8
Interaction
Interacts with ATG3, ATG8, ATG12, EP300 and FOXO1
Cell Type
Mature Neurons, Neurons
Biology Area
Apoptosis/Tumor Suppressors/Cell Death, Cell Biology, Cell Death, Cell Proliferation and Viability, Neurodegeneration, Protein Misfolding and Aggregation, Protein Trafficking and Clearance, Ubiquitin/Protein Degradation
Molecular Family
Autophagosome Markers, Ligases
Antigen References
  1. Komatsu M, et al. 2005. J Cell Biol. 169:425-34.
  2. Komatsu M, et al. 2007. Proc Natl Acad Sci USA. 104:14489-94.
  3. Lee I H, et al. 2012. Science. 336: 225–228.
  4. Collier JJ, et al. 2021. EMBO Mol Med. 13:e14824.
Gene ID
10533 View all products for this Gene ID
UniProt
View information about ATG7 on UniProt.org
Go To Top Version: 1    Revision Date: 10/17/2023

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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