ATG8 polyclonal antibody
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More Files
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Western Blot (Recombinant protein)
Immunoblot of ATG8 fusion protein.
ATG8 polyclonal antibody (Cat # PAB11359) generated by immunization with recombinant yeast ATG8 was tested by immunoblot with other anti-UBL antibodies against E.coli lysates expressing the ATG8-GFP fusion protein.
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 APG8 using a 1:4,000 and 1:8,000 dilution of ATG8 polyclonal antibody (Cat # PAB11359) (Yeast) followed by reaction with a 1:15,000 dilution of HRP Goat-a-Rabbit IgG MX.
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.
This data indicates that anti-ATG8 is highly specific and does not cross react with other UBLs. A chemiluminescence system was used for signal detection (Roche) .
Data contributed by M. Malakhov, www.lifesensors.com, personal communication.
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Specification
Product Description
Rabbit polyclonal antibody raised against recombinant ATG8.
Immunogen
Recombinant protein corresponding to Saccharomyces cerevisiae ATG8.
Host
Rabbit
Reactivity
Yeast
Specificity
This is an IgG fraction antibody 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.
Form
Lyophilized
Recommend Usage
ELISA (1:20000-1:100000)
Western Blot (1:4000-1:8000)
The optimal working dilution should be determined by the end user.Storage Buffer
Lyophilized from 20 mM potassium phosphate buffer, 150 mM NaCl, pH 7.2 (0.01% sodium azide).
Storage Instruction
Store at 4°C prior to restoration. After reconstitution with 100 uL of deionized water (or equivalent), store at -20°C or below for long term storage.
Aliquot to avoid repeated freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4°C as an undiluted liquid. Dilute only prior to immediate use.Note
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. For immunoblotting a 14 kDa band corresponding to yeast APG8 is detected. Most yeast cell lysates can be used as a positive control without induction or stimulation.
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only. -
Applications
Western Blot (Recombinant protein)
Immunoblot of ATG8 fusion protein.
ATG8 polyclonal antibody (Cat # PAB11359) generated by immunization with recombinant yeast ATG8 was tested by immunoblot with other anti-UBL antibodies against E.coli lysates expressing the ATG8-GFP fusion protein.
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 APG8 using a 1:4,000 and 1:8,000 dilution of ATG8 polyclonal antibody (Cat # PAB11359) (Yeast) followed by reaction with a 1:15,000 dilution of HRP Goat-a-Rabbit IgG MX.
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.
This data indicates that anti-ATG8 is highly specific and does not cross react with other UBLs. A chemiluminescence system was used for signal detection (Roche) .
Data contributed by M. Malakhov, www.lifesensors.com, personal communication.Enzyme-linked Immunoabsorbent Assay
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Gene Info — ATG8
Entrez GeneID
852200Protein Accession#
NP_009475; P38182Gene Name
ATG8
Gene Alias
APG8, AUT7, CVT5
Gene Description
Atg8p
Gene Ontology
HyperlinkGene Summary
Atg7p
Other Designations
Protein required for autophagy; modified by the serial action of Atg4p, Atg7p, and Atg3p, and conjugated at the C terminus with phosphatidylethanolamine, to become the form essential for generation of autophagosomes
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Publication Reference
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Human autophagins, a family of cysteine proteinases potentially implicated in cell degradation by autophagy.
Marino G, Uria JA, Puente XS, Quesada V, Bordallo J, Lopez-Otin C.
The Journal of Biological Chemistry 2002 Nov; 278(6):3671.
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Molecular dissection of autophagy: two ubiquitin-like systems.
Ohsumi Y.
Molecular and Cellular Biology 2001 Mar; 2(3):211.
Application：WB-Tr, Yeast, Yeast cells.
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Ubiquitin and its kin: how close are the family ties?
Jentsch S, Pyrowolakis G.
Trends in Cell Biology 2000 Aug; 10(8):335.
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Human autophagins, a family of cysteine proteinases potentially implicated in cell degradation by autophagy.
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