Anti-ATG8 antibody (ab4753)
Key features and details
- Rabbit polyclonal to ATG8
- Suitable for: WB, ELISA
- Reacts with: Saccharomyces cerevisiae
- Isotype: IgG
Overview
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Product name
Anti-ATG8 antibody -
Description
Rabbit polyclonal to ATG8 -
Host species
Rabbit -
Tested applications
Suitable for: WB, ELISAmore details -
Species reactivity
Reacts with: Saccharomyces cerevisiae -
Immunogen
Recombinant full length protein (S. cerevisiae).
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped 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 buffer
pH: 6.50
Preservative: 0.01% Sodium azide
Constituents: 0.42% Potassium phosphate, 0.87% Sodium chloride -
Concentration information loading...
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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
Associated products
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
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.
Application | Abreviews | Notes |
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WB | (4) |
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.
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ELISA |
1/20000 - 1/100000.
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Notes |
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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. |
ELISA
1/20000 - 1/100000. |
Target
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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 -
Database links
- Entrez Gene: 852200 Saccharomyces cerevisiae
- SwissProt: P38182 Saccharomyces cerevisiae
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Alternative names
- Apg 8 antibody
- APG8 antibody
- Atg8p antibody
see all
Images
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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
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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.
Datasheets and documents
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SDS download
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Datasheet download
References (10)
ab4753 has been referenced in 10 publications.
- Dabin R et al. Astrocytic IGF-1 and IGF-1R Orchestrate Mitophagy in Traumatic Brain Injury via Exosomal miR-let-7e. Oxid Med Cell Longev 2022:3504279 (2022). PubMed: 36062186
- Calero-Muñoz N et al. Cadmium induces reactive oxygen species-dependent pexophagy in Arabidopsis leaves. Plant Cell Environ 42:2696-2714 (2019). PubMed: 31152467
- Li X et al. Dehydrin MtCAS31 promotes autophagic degradation under drought stress. Autophagy N/A:1-16 (2019). PubMed: 31362589
- 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 . 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). PubMed: 28596290
- Spencer B et al. a-Synuclein interferes with the ESCRT-III complex contributing to the pathogenesis of Lewy body disease. Hum Mol Genet 25:1100-15 (2016). PubMed: 26740557
- Shemi A et al. Phosphorus starvation induces membrane remodeling and recycling in Emiliania huxleyi. New Phytol 211:886-98 (2016). PubMed: 27111716
- Li S et al. Expression of Malus xiaojinensis IRT1 (MxIRT1) protein in transgenic yeast cells leads to degradation through autophagy in the presence of excessive iron. Yeast 32:499-517 (2015). WB . PubMed: 25871543
- Schatz D et al. Hijacking of an autophagy-like process is critical for the life cycle of a DNA virus infecting oceanic algal blooms. New Phytol 204:854-63 (2014). WB, Electron Microscopy . PubMed: 25195618
- Crews L et 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 . PubMed: 20174468