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Products:Cell Biology >> Proteolysis / Ubiquitin >> Proteasome / Ubiquitin >> Ub-like Proteins
Overview
Product name
Anti-ATG8 antibody
Description
Rabbit polyclonal to ATG8
Tested applications
WB, IHC-P, ELISAmore details
Cross reactivity
Reacts with
Saccharomyces cerevisiae
Immunogen
Recombinant full length protein (S. cerevisiae).
Properties
Form
Liquid
Storage instructions
Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage buffer
Preservative: 0.01% Sodium Azide
Constituents: 0.15M Sodium Chloride, 0.02M Potassium Phosphate. pH 7.2
Concentration
Concentration information loading...
Purity
IgG fraction
Purification notes
Purified 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.
Clonality
Polyclonal
Isotype
IgG
Research areas
Cardiovascular >> Heart >> Autophagy >> APG gene products
Microbiology >> Organism >> Eukaryotic Microorganism >> Yeast >> Saccharomyces spp
Neuroscience >> Neurology process >> Neurodegenerative disease >> Other
Signal Transduction >> Protein Trafficking >> Vesicle Transport >> SNAPs & SNAREs
Cell Biology >> Proteolysis / Ubiquitin >> Proteasome / Ubiquitin >> Ub-like Proteins
Applications
Show applications key
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 dilution.
ELISA: 1/20000 - 1/100000.
Target
Relevance
ATG8 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.
Cellular localization
Cytoplasmic
Alternative names
- Apg 8 antibody
- APG8 antibody
- Atg8p antibody
see all
Database links
- Entrez Gene: 852200 Saccharomyces cerevisiae
- SwissProt: P38182 Saccharomyces cerevisiae
Anti-ATG8 antibody images:
Western blot - ATG8 antibody (ab4753)
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 reactivity of anti-APG8 (anti-Apg 8) with other UBLs. This data indicates that anti-ATG8 (anti-Apg8)is highly specific and does not cross react with other UBLs. A chemiluminescence system was used for signal detection (Roche). Other detection systems will yield similar results.
Data contributed by M. Malakhov, www.lifesensors.com, personal communication.
Western blot - ATG8 antibody (ab4753)
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
Secondary
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.
This image is courtesy of an anonymous abreview.
References for Anti-ATG8 antibody (ab4753)
This product has been referenced in:
- Crews Let al. Selective Molecular Alterations in the Autophagy Pathway in Patients with Lewy Body Disease and in Models of alpha-Synucleinopathy. PLoS One 5:e9313 (2010). WB, IHC-P; Human, Mouse.Read more (PubMed: 20174468) »
See 1 publication for this product
Publishing research using ab4753? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"