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.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG₁

Scientific Data Images for Human/Mouse ATG7 Antibody

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.

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 Northern-Lights™ 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.

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 counter-stained 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.

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 Western^TM.

Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for ATG7 at approximately 72 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.

Detection of Human ATG7 by Western Blot

ATG5 and ATG7 enhanced autophagy and inhibited ER stress in chondrocyte. a Western blotting analysis of LC3, P62, ATG5, ATG7 and ATG5-ATG12 expression after infected with Ad-ATG5, Ad-ATG7 and Ad-ATG5 + Ad-ATG7 in the C28I2 cells. beta-actin is served as an internal control. b Qualitative analysis of ATG5, ATG7, ATG5-ATG12, LC3 and P62. The values were normalized to beta-actin. c C28I2 cells were double stained with LC3 (red) and DAPI (blue) and visualized by confocal microscopy (400X) after treated with Rapamycine, Ad-ATG5, Ad-ATG7 and Ad-ATG5 + Ad-ATG7 24 h. *P < 0.05, **P < 0.01 compared with the controls. Values are means ± SD n = 3). d Qualitative analysis of LC3 fluorescence intensity of chondrocytes. The values were normalized to the NC group. e Western blotting analysis of PERK, p-PERK and Nrf2 expression after infected with Ad-ATG5, Ad-ATG7 and Ad-ATG5 + Ad-ATG7 in the C28I2 cells. beta-actin is served as an internal control. f Qualitative analysis of PERK, p-PERK and Nrf2 were normalized to beta-actin. (1:NC, 2:Ad-GFP, 3:Ad-ATG5, 4:RAPA, 5:Ad-ATG7, 6:Ad-ATG5 + RAPA, 7:Ad-ATG5 + Ad-ATG7). Rapamycin (25 μM) used as a positive control Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31060556), licensed under a CC-BY license. Not internally tested by R&D Systems.

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Applications for Human/Mouse ATG7 Antibody

Application

Recommended Usage

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
Sample: Immersion fixed RAW 264.7 mouse monocyte/macrophage cell line

Immunohistochemistry

8-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human brain (cortex)

Intracellular Staining by Flow Cytometry

0.25 µg/10⁶ cells
Sample: HeLa human cervical epithelial carcinoma cell line fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005)

Simple Western

20 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line

Western Blot

2 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line and HepG2 human hepatocellular carcinoma cell line

Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 3 reviews rated 5 using MAB6608 in the following applications:

Immunohistochemistry (2 Reviews)
Western Blot (1 Review)

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Sterile PBS to a final concentration of 0.5 mg/mL. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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

Alternate Names

APG7, APG7L, GSA7

Entrez Gene IDs

10533 (Human); 74244 (Mouse); 312647 (Rat); 856576 (Yeast)

Gene Symbol

ATG7

UniProt

O95352

Additional ATG7 Products

Product Documents for Human/Mouse ATG7 Antibody

Product Datasheets

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COA

SDS

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Product Specific Notices for Human/Mouse ATG7 Antibody

For research use only

Citations
Reviews

Calculators
Flow Cytometry

Human/Mouse ATG7 Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB6608

Key Product Details

Validated by

Species Reactivity

Validated:

Cited:

Applications