Western Blot: ATG5 Antibody [NB110-53818] - Total protein from Human HeLa and A431 and Mouse MEF cells was separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The ...read more
Immunocytochemistry/ Immunofluorescence: ATG5 Antibody [NB110-53818] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.05% Triton-X100. The cells were ...read more
Immunohistochemistry: ATG5 Antibody [NB110-53818] - Staining of human liver hepatocytes at 2.5ug/ml. 40X magnification.
Western Blot: ATG5 Antibody [NB110-53818] - Analysis in mouse wildtype ES cell lysate (Lane 1) using NB110-53818. Lane 2 is a mouse ATG5 KO ES cell lysate (negative control). Atg5-/- ES cells from Dr. Noboru Mizushima ...read more
Immunocytochemistry/ Immunofluorescence: ATG5 Antibody [NB110-53818] - Staining of SY5Y cells at 1:250. Incubated overnight at 4 degrees. Photo courtesy of an anonymous collaborator.
Immunocytochemistry/ Immunofluorescence: ATG5 Antibody [NB110-53818] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. The cells were ...read more
Immunohistochemistry: ATG5 Antibody [NB110-53818] - Staining mouse intestine using DAB with hematoxylin counterstain.
Immunohistochemistry-Paraffin: ATG5 Antibody [NB110-53818] - Staining of mouse knee. Image from 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
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).
In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
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-like (S. cerevisiae)
ATG5 autophagy related 5 homolog (S. cerevisiae)
Autophagy protein 5
ATG5 (autophagy protein 5, also known as APG5-like, apoptosis-specific protein, APG5L, and ASP) belongs to ATG5 family and is an essential protein for autophagy. It is expressed ubiquitously and the expression increases dramatically in apoptotic cells. ATG5 is localized in cytoplasm (can colocalize with nonmuscle actin) and is induced upon apoptotic stimuli. ATG5-ATG12 conjugate forms a complex with several units of ATG16 and this complex inhibits ATG5-TECPR1 direct interactions. After conjugating to ATG12, ATG5 associates with the isolation membrane to form autophagosomes and this conjugation reaction is mediated by ubiquitin E1-like enzyme Atg7 as well as the E2-like enzyme Atg10. The conjugate detaches from the membrane immediately before or after the completion of autophagosome formation. ATG5 play an important role in the apoptotic process, within the modified cytoskeleton, and its expression is a relatively late event in apoptotic process, occurring downstream of caspase activity.
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.
Karanasios E, Ktistakis NT. Studying Autophagy: List of Useful Antibodies Produced via a Community Effort. Autophagy at the Cell, Tissue and Organismal Level. 2016 Apr 24
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)
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 Immunofluorescence, 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.
Could you help me with the following question about ATG5 antibody (Cat. NB110-53818) that I bought from you? The arrow in the data sheet points to Atg5/Atg12 band of 56kDa, there is also a band of 33kDa which I guess is the Atg5? Could you excuse my ignorance and explain what is the meaning of detecting the Atg5/Atg12 conjugate in preference to Atg5, I'm obviously missing something and Wikipedia has nothing on the subject. Do I guess correctly that the antibody does not 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 bonded with Atg12, this is a ubiquitous reaction. The lysate that was used was meant to illustrate the ability to detect autophagy by WB via Atg5/Atg12. It is possible to detect free floating 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 is not able to detect Atg12, which free floating runs at about 23 kDa.
As you mention in 3rd FAQs (Frequently asked questions) section, that this antibody can detect free ATG5 in cell lysates (although there is no western blot image). Does this antibody can detect free ATG5 monomer in Hela or HEK 293 cells?
With regard to our product NB110-53818 and the FAQ #3, this antibody is able to detect the ATG5 (unconjugated form) ~33kDa and the conjugated form ~56kDa. In the western blot image we have on our website, we show the antibody's ability to detect the autophagy induced conjugate ATG5/ATG12. We also have in a customer review, an image of both the conjugate and unconjugated ATG5 in rat Cardiomyocyte. We also have quite a few publications that show this antibody working in western blot with mouse and human samples. I am not sure if any of the publications have specifically tested HeLa or HEK 293 cells, but if the ATG5 protein is present, this product will be able to 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 samples.