The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration. PubMed: 21784844
Use a concentration of 10 µg/ml.
Use a concentration of 0.5 - 1 µg/ml. Detects a band of approximately 78 kDa (predicted molecular weight: 78 kDa).
Use at an assay dependent concentration. PubMed: 20174468
E1-like activating enzyme involved in the 2 ubiquitin-like systems required for cytoplasm to vacuole transport (Cvt) and autophagy. Activates ATG12 for its conjugation with ATG5 as well as the ATG8 family proteins for their conjugation with phosphatidylethanolamine. Both systems are needed for the ATG8 association to Cvt vesicles and autophagosomes membranes. Required for autophagic death induced by caspase-8 inhibition. Required for mitophagy which contributes to regulate mitochondrial quantity and quality by eliminating the mitochondria to a basal level to fulfill cellular energy requirements and preventing excess ROS production. Modulates p53/TP53 activity to regulate cell cycle and survival during metabolic stress. Plays also a key role in the maintenance of axonal homeostasis, the prevention of axonal degeneration, the maintenance of hematopoietic stem cells, the formation of Paneth cell granules, as well as in adipose differentiation.
Widely expressed, especially in kidney, liver, lymph nodes and bone marrow.
Belongs to the ATG7 family.
The C-terminal part of the protein is essential for the dimerization and interaction with ATG3 and ATG12. The N-terminal FAP motif (residues 15 to 17) is essential for the formation of the ATG89-PE and ATG5-ATG12 conjugates.
Acetylated by EP300.
Cytoplasm. Preautophagosomal structure. Localizes also to discrete punctae along the ciliary axoneme and to the base of the ciliary axoneme.
IHC image of ab53255 staining in human breast carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab53255, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-Apg7 antibody (ab53255)Image from Meng Q & Cai D, J Biol Chem. 2011 Sep 16;286(37):32324-32. Epub 2011 Jul 22. Fig 4.; doi: 10.1074/jbc.M111.254417; September 16, 2011, The Journal of Biological Chemistry, 286, 32324-32332.
Immunohistochemical analysis of murine brain tissue, staining Apg7 with ab53255.
Fixed brain sections were blocked with serum, permeabilized with 0.2% Triton X-100, and treated with primary antibody. An AlexaFluor®555-conjugated anti-rabbit IgG was used as the secondary antibody.
Sun S et al. ATG7 promotes the tumorigenesis of lung cancer but might be dispensable for prognosis predication: a clinicopathologic study. Onco Targets Ther9:4975-81 (2016).
Read more (PubMed: 27563251) »
Liu L et al. Baclofen mediates neuroprotection on hippocampal CA1 pyramidal cells through the regulation of autophagy under chronic cerebral hypoperfusion. Sci Rep5:14474 (2015).
Read more (PubMed: 26412641) »