Images
Western Blot (Recombinant protein)
Application

Western Blot (Recombinant protein)

Immunoblot of APG12 fusion protein.
ATG12 polyclonal antibody (Cat # PAB11358) generated by immunization with recombinant yeast ATG12 was tested by immunoblot against yeast lysates expressing the ATG12-GFP fusion protein and other UBL fusion proteins.
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 positions of free GFP or GFP containing recombinant proteins present in each lysate preparation after reaction with a 1:1,000 dilution of anti-GFP followed by reaction with a 1:15,000 dilution of HRP Donkey-a-Goat IgG MX.
Panel C shows specific reaction with ATG12 using a 1:2,000 dilution of ATG12 polyclonal antibody (Cat # PAB11358) 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.
Yeast lysate proteins were separated by SDS-PAGE using a 15% gel. This data indicates that anti-ATG12 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, Lifesensors Inc., personal communication.

  • Specifications

    Product Description

    Rabbit polyclonal antibody raised against recombinant ATG12.

    Immunogen

    Recombinant protein corresponding to Saccharomyces cerevisiae ATG12.

    Host

    Rabbit

    Reactivity

    Yeast

    Form

    Lyophilized

    Quality Control Testing

    Antibody Reactive Against Recombinant Protein.

    Recommend Usage

    ELISA (1:1000-1:5000)
    Western Blot (1:2000)
    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. For long term storage store at -20°C.
    Aliquot after reconstitution to avoid repeated freezing and thawing.

    Note

    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 APG12 fusion protein.
    ATG12 polyclonal antibody (Cat # PAB11358) generated by immunization with recombinant yeast ATG12 was tested by immunoblot against yeast lysates expressing the ATG12-GFP fusion protein and other UBL fusion proteins.
    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 positions of free GFP or GFP containing recombinant proteins present in each lysate preparation after reaction with a 1:1,000 dilution of anti-GFP followed by reaction with a 1:15,000 dilution of HRP Donkey-a-Goat IgG MX.
    Panel C shows specific reaction with ATG12 using a 1:2,000 dilution of ATG12 polyclonal antibody (Cat # PAB11358) 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.
    Yeast lysate proteins were separated by SDS-PAGE using a 15% gel. This data indicates that anti-ATG12 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, Lifesensors Inc., personal communication.

    Enzyme-linked Immunoabsorbent Assay

  • Gene Info — ATG12

    Entrez GeneID

    852518

    Gene Name

    ATG12

    Gene Alias

    APG12

    Gene Description

    Atg12p

    Gene Ontology

    Hyperlink

    Gene Summary

    conjugated via an isopeptide bond to a lysine residue of Atg5p by the E1 enzyme

    Other Designations

    Ubiquitin-like modifier, conjugated via an isopeptide bond to a lysine residue of Atg5p by the E1 enzyme, Atg7p, and the E2 enzyme, Atg10p, a step that is essential for autophagy

  • Publication Reference