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.
Datasheet:
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Publication Reference
1.
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.J Biol Chem. 2003 Feb 7;278(6):3671-8. Epub 2002 Nov 21.
2.
Molecular dissection of autophagy: two ubiquitin-like systems.Ohsumi Y.Nat Rev Mol Cell Biol. 2001 Mar;2(3):211-6.
3.
Ubiquitin and its kin: how close are the family ties?Jentsch S, Pyrowolakis G.Trends Cell Biol. 2000 Aug;10(8):335-42.
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
Application Image
Western Blot (Recombinant protein)
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Enzyme-linked Immunoabsorbent Assay
Gene Information
Entrez GeneID:
852200
Protein Accession#:
NP_009475; P38182
Gene Name:
ATG8
Gene Alias:
APG8,AUT7,CVT5
Gene Description:
Atg8p
Gene Ontology:
Hyperlink
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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