Genetic Strategies: Western Blot: ATG5 Antibody [NB110-53818] - Western blot analysis in mouse wildtype ES cell lysate (Lane 1) using NB110-53818. Lane 2 is a mouse ATG5 KO ES cell lysate (negative control). ...read more
Genetic Strategies: Western Blot: ATG5 Antibody [NB110-53818] - ERK phosphorylation in Atg5-/-cells depends on nutrient availability. Serum-deprived Atg5-/- MEFs display decreased ERK and p90RSK phosphorylation. ...read more
Immunocytochemistry/ Immunofluorescence: ATG5 Antibody [NB110-53818] - ICC/IF analysis of SY5Y cells at 1:250. Incubated overnight at 4 degrees. Photo courtesy of an anonymous collaborator.
Immunohistochemistry: ATG5 Antibody [NB110-53818] - Immunohistochemistry analysis of human liver hepatocytes at 2.5 ug/mL. 40X magnification.
Immunocytochemistry/ Immunofluorescence: ATG5 Antibody [NB110-53818] - ERK2 utilizes kinase-docking domains to interact with ATG5-ATG12 and LC3-II.(a) Mutations in FRS on ERK2 decrease colocalization of ERK2 with ...read more
Western Blot: ATG5 Antibody [NB110-53818] - Western blot analysis of total protein from Human HeLa and A431 and Mouse MEF cells. Lysates were separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked ...read more
Biological Strategies: Western Blot: ATG5 Antibody [NB110-53818] - IFN-gamma increased rates of autophagy in primary BMECs in vitro. BMECs were treated with IFN-gamma at 2.5, 5, 10 or 20 ng/mL. At the end of the ...read more
Immunohistochemistry: ATG5 Antibody [NB110-53818] - Immunohistochemistry analysis of mouse intestine using DAB with hematoxylin counterstain.
Immunohistochemistry-Paraffin: ATG5 Antibody [NB110-53818] - Immunohistochemistry analysis of mouse knee. IHC-P image submitted by a verified customer review.
Simple Western: ATG5 Antibody [NB110-53818] - Simple Western lane view shows a specific band for ATG5 in 0.5 mg/mL of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation ...read more
Genetic Strategies: Knockdown Validated: ATG5 Antibody [NB110-53818] - Knockdown of autophagy-related gene 5 (ATG5) inhibits trehalose-induced autophagy in normal human primary airway epithelial cells. Normal ...read more
In Western Blot, a band is seen ~56 kDa representing the ATG5-ATG12 complex, the molecular weight of human ATG5 is ~33 kDa. In ICC/IF, cytoplasmic staining was observed in SY5Y cells. In IHC-P, staining was observed in the cytoplasm of human hepatocytes and mouse intestine tissues. Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (pH 6.0) is recommended. Use in Immunoprecipitation reported in scientific literature (PMID: 24705551). Use in radioimmunoassay reported in scientific literature (PMID: 28651493). PLA data from independent lab. Use in electron microscopy reported in scientific literature (PMID: 27219062). Use in immunoblotting reported in scientific literature (PMID: 23918802). Use in ELISA reported in scientific literature (PMID: 25484078). Use in electron microscopy reported in scientific literature (PMID: 27219062). Use in ICC/IF was reported in scientific literature (PMID: 18073215).
In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
32 kDa. Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Fish reactivity reported in scientific literature (PMID: 26183773). Guinea Pig reactivity reported in scientific literature (PMID: 30766882).
Packaging, Storage & Formulations
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
0.02% Sodium Azide
Immunogen affinity purified
Alternate Names for ATG5 Antibody
APG5 (Autophagy 5, S. Cerevisiae)-Like
APG5 autophagy 5-like (S. cerevisiae)
ATG5 autophagy related 5 homolog (S. cerevisiae)
ATG5 Autophagy Related 5 Homolog
Autophagy protein 5
Autophagy Related 5
Atg5, Autophagy related 5 or autophagy protein 5 (theoretical molecular weight 32 kDa), belongs to a group of core autophagy-related proteins first identified in yeast and later in eukaryotic cells. Atg proteins play essential roles in the process or macroautophagy. Atg5 is considered a core autophagy protein, for its role in the formation of the autophagosome, a double membrane vesicle which engulfs proteins and organelles for delivery to the lysosome and subsequent degradation (1). Atg5 participates in the process of phagophore elongation by interacting with the ubiquitin-like protein Atg12. Formation of the Atg12-Atg5 conjugate is dependent on the activities of Atg7 (E1 ubiquitin-activating like enzyme) and Atg10 (E2 ubiquitin-activating like enzyme). Non-covalent interaction between the Atg12-Atg5 conjugate and Atg16L1, allows for the formation of a large complex which associates with the nascent phagophore. The Atg16L1 complex dissociates from the autophagosome once it is fully formed (1,2).
In the context of its role in autophagy, Atg5 plays diverse physiologically relevant roles. For example, Atg5 together with Atg7 are required for adipogenesis (3). Recently, Atg5 has been implicated in the process of B-cell receptor polarization and antigen presentation (4). In addition to its role in autophagy, Atg5 is implicated in apoptotic cell death. Interaction of Atg5 with FADD (Fas-associated protein with death domain) is involved in cell death induced by IFN-gamma. A truncated form of Atg5, a 24kDa fragment, leads to cell death by interacting with Bcl-xl and inhibiting its anti-apoptotic activity (5). Other Atg5 interacting partners include interleukin-beta (IL-beta) converting enzyme and nucleotide binding oligomerization domain protein 1, which suggest that Atg5 may play other biologically relevant roles (3).
1. Yang, Z., & Klionsky, D. J. (2010). Mammalian autophagy: Core molecular machinery and signaling regulation. Current Opinion in Cell Biology. https://doi.org/10.1016/j.ceb.2009.11.014
2. Rubinsztein, D. C., Shpilka, T., & Elazar, Z. (2012). Mechanisms of autophagosome biogenesis. Current Biology. https://doi.org/10.1016/j.cub.2011.11.034
3. Subramani, S., & Malhotra, V. (2013). Non-autophagic roles of autophagy-related proteins. EMBO Reports. https://doi.org/10.1038/embor.2012.220
4. Arbogast, F., Arnold, J., Hammann, P., Kuhn, L., Chicher, J., Murera, D., Gros, F. (2019). ATG5 is required for B cell polarization and presentation of particulate antigens. Autophagy. https://doi.org/10.1080/15548627.2018.1516327
5. Luo, S., & Rubinsztein, D. C. (2007). Atg5 and Bcl-2 provide novel insights into the interplay between apoptosis and autophagy. Cell Death and Differentiation. https://doi.org/10.1038/sj.cdd.4402149
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.
Xia X, Gao Y, Zhang J et al. Autophagy mediated by arginine depletion activation of the nutrient sensor GCN2 contributes to interferon-gamma-induced malignant transformation of primary bovine mammary epithelial cells Cell Death Discovery Jan 25 2016 (WB, Bovine)
Ruhen O Comprehensive genomic profiling of circulating tumour DNA and tumour-derived extracellular vesicles from breast cancer patients Thesis (WB, Human)
Wible D, Chao H, Tang D, Bratton S ATG5 cancer mutations and alternative mRNA splicing reveal a conjugation switch that regulates ATG12 ATG5-ATG16L1 complex assembly and autophagy Cell Discov Dec 1 2019 (WB, Human)
FAQs for ATG5 Antibody (NB110-53818). (Showing 1 - 5 of 5 FAQs).
I am interested in testing your anti ATG5 (NB110-53818SS) for Immunofluorescence in MDCK cell (canine epithelial cell line). I noticed you report reactivity of the antibody against several species but not dog. Do you know if the Ab will react against canine genome? Also for IF what fixation do you recommend?
The applications and species listed for any one of our antibodies are ones we have tested and validated. Canine is not listed as it has not been tested and therefore we cannot guarantee its cross reactivity. If you would be interested in testing this antibody in canine, we can recommend our Innovator's Reward. Our Innovator's Reward is offered to reward researchers for testing new species and applications with our products. Learn more about our Innovators Reward Program. Also, please use this link to view our Immunofluorescence protocol which explains the fixation method that we recommend.
Our customer is interested in this ATG5 antibody (cat. no. NB110-53818). Would you please confirm that this antibody would recognize ATG5 alone, as well as ATG5-ATG12 conjugate?
The antibody was raised against a peptide corresponding to the N-terminal of ATG5. It should recognize ATG5 alone but it appears that ATG5 is rarely found alone in the cell.
The arrow in the data sheet points to the ATG5/ATG12 band of 56 kDa. There is also a band of 33 kDa which I guess is ATG5? Could you explain the meaning of detecting the ATG5/ATG12 conjugate in preference to ATG5? Does the antibody cross-react with ATG12?
The Western blot image you are referring to illustrates the ATG5/ATG12 conjugate, which is formed during autophagy. During autophagy ATG5 is covalently bonds ATG12 in a ubiquitous reaction. The lysate that was used was meant to illustrate the ability to detect autophagy by Western blot via ATG5/ATG12. It is possible to detect free ATG5 in cell lysates, although it is not seen in this Western blot. The lower 2 bands do not correspond to ATG5. Our ATG5 antibody does not detect ATG12, which runs at about 23 kDa.
Does this antibody detect free ATG5 monomer in HeLa or HEK 293 cells?
This antibody is able to detect ATG5 (unconjugated form) at ~33 kDa and the ATG12-conjugated form at ~56 kDa. In the Western blot image we have on our website, we show the antibody's ability to detect the autophagy induced conjugate ATG5/ATG12. A customer review image shows both the conjugated and unconjugated ATG5 in rat cardiomyocytes. Several publications show this antibody working in Western blot with mouse and human samples. We are not sure if in any of the publications HeLa or HEK 293 cells specifically were used, but if the ATG5 protein is present, this product will detect it in Western blot.
Would you please help confirm if NB110-53818 has been tested by IHC with frozen sections?
NB110-53818 has been tested with paraffin-embedded tissue, but we have not validated this antibody for IHC with frozen tissue sections.
ATG9A - early marker autophagosome assembly ATG9A is the only essential integral membrane protein involved in autophagy. ATG9A contains six transmembrane domains and initiates the assembly of autophagosomes. The autophagosome is a double-membrane structure that engulfs and eventually degrade... Read full blog post.
ATG5: Roles in Cellular Defense ATG5, or Autophagy Related 5, is a protein crucial for autophagy. Autophagy is a mechanism in which dysfunctional or pathogenic cells or cellular components are degraded and sometimes recycled. This process happens when ATG5 conjugates with another pr... Read full blog post.
ATG5, Autophagy and Apoptosis ATG5 is a member of the ATG family that regulates autophagy, the evolutionary conserved homeostatic response to a diverse variety of self- and foreign-originating cellular stresses. ATG5 is ubiquitously expressed in cells and found co-localized with c... Read full blog post.
ATG5: From Autophagy to Alzheimer's Disease Autophagy is a conserved mechanism whereby cells form double membrane autophagosomes to sequester cytoplasmic components for subsequent destruction by fusion with lysosomes (eukaryotes) or vacuoles (yeast). Targets of autophagy include aging proteins,... Read full blog post.
Analyzing LC3 in Western blot Microtubule-associated protein light chain 3 (LC3) is considered one of the definitive markers of autophagy, and its use is widespread in labs throughout the world. Despite its popularity, there are several considerations when employing LC3 antibodies... Read full blog post.
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PRODUCT AVAILABILITY: Update Regarding the Evolving COVID-19 Situation
Bio-Techne appreciates the critical role that you and our products and services play in research efforts to further scientific innovation and discovery. We are continually assessing our manufacturing and supplier capabilities during the COVID-19 situation and are implementing precautionary measures to ensure uninterrupted supply of products and services. Currently, and as we abide by local shelter in place orders across the world, we are fully operational and do not anticipate any material supply disruptions across our Bio-Techne brands and product lines. As the situation evolves, our goal is to utilize preventive measures to reduce the threat that COVID-19 poses to our ability to meet the needs of our customers globally.