Product nameAnti-ATG8 antibody
DescriptionRabbit polyclonal to ATG8
Tested applicationsSuitable for: WB, IHC-P, ELISA, Electron Microscopymore details
Species reactivityReacts with: Saccharomyces cerevisiae
Recombinant full length protein (S. cerevisiae).
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 0.42% Potassium phosphate, 0.87% Sodium chloride
Concentration information loading...
Purification notesPurified from monospecific antiserum by a multi-step process which includes delipidation, salt fractionation and ion exchange chromatography followed by extensive dialysis against the buffer stated above. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-rabbit serum.
Our Abpromise guarantee covers the use of ab4753 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/4000 - 1/8000. Detects a band of approximately 14 kDa. This antibody using the specified conditions may recognize other prominent intrinsic bands (UBLs or their conjugates). Other intrinsic bands are readily detectable in yeast lysates at lower antibody dilutions.|
|IHC-P||Use at an assay dependent concentration. PubMed: 20174468|
|ELISA||1/20000 - 1/100000.|
|Electron Microscopy||Use at an assay dependent concentration. PubMed: 25195618|
RelevanceATG8 is a ubiquitin-like protein in yeast required for autophagy (intracellular bulk protein degradation). Starved yeast cells take up their own cytoplasm into vacuoles through autophagic bodies. ATG8 is requiered for autophagy: modified by the serial action of ATG4, ATG7, and ATG3, and conjugated at the C terminus with phosphatidylethanolamine, to become the form essential for generation of autophagosomes. ATG8 interacts also with the endoplasmic reticulum to Golgi v-SNARE protein BET1 and the vacuolar v-SNARE protein NYV1. ATG8 is highly conserved, with apparent homologues in the worm, mammals and plants. In higher eukaryotes, ATG8 consists of a multigene family.
- Apg 8 antibody
- APG8 antibody
- Atg8p antibody
Ab4753, generated by immunization with recombinant yeast ATG8 (or Apg 8), was tested by immunoblot with other anti-UBL (Ubiquitin-like modifier) antibodies against E.coli lysates expressing the ATG8-GFP fusion protein (Apg 8-GFP in both panels)). All UBLs possess limited homology to Ubiquitin and to each other, therefore it is important to know the degree of reactivity of each antibody against each UBL.
Panel A shows total protein staining using ponceau.
Panel B shows specific reaction with ATG8 (Apg 8) using a 1:4,000 and 1:8,000 dilution of ab4753 followed by reaction with a 1:15,000 dilution of HRP Goat-a-Rabbit IgG (ab7090). All primary antibodies were diluted in TTBS buffer supplemented with 5% non-fat milk and incubated with the membranes overnight at 4° C. E.coli lysate proteins were separated by SDS-PAGE using a 15% gel.
Similar experiments (data not shown), where other UBL fusion proteins were separated and probed with this antibody showed no react
Anti-ATG8 antibody (ab4753) at 1/1500 dilution + Whole cell lysate prepared from Human huh-7 cells expressing the Saccharomyces cerevisiae protein at 15 µg
Sheep anti-rabbit IgG conjugated to HRP at 1/7000 dilution
Observed band size: 15 kDa why is the actual band size different from the predicted?
Exposure time: 4 minutes
Gel run under denaturing conditions.Primary antibody incubated for 10 minutes at 20°C.Blocked using 0.5% milk for 1 minute at 20°C.Detection method: Western lightning chemiluminescent reagent.
Immuno-TEM using ab4753 at 1/30 dilution to label purified virions of a DNA coccolithovirus.
Data from: New Phytol. 2014 Dec;204(4):854-63 (PMID 25195618)
Daniella Schatz, Adva Shemi, Shilo Rosenwasser, Helena Sabanay, Sharon G Wolf, Shifra Ben-Dor, and Assaf Vardi
This product has been referenced in:
- Zhou S et al. Autophagy contributes to regulate the ROS levels and PCD progress in TMV-infected tomatoes. Plant Sci 269:12-19 (2018). WB . Read more (PubMed: 29606209) »
- Jo M et al. Yeast genetic interaction screen of human genes associated with amyotrophic lateral sclerosis: identification of MAP2K5 kinase as a potential drug target. Genome Res 27:1487-1500 (2017). Read more (PubMed: 28596290) »