Images
Western Blot (Recombinant protein)
Application

Western Blot (Recombinant protein)

Immunoblot of HUB1 fusion protein.
HUB1 polyclonal antibody (Cat # PAB11361) generated by immunization with recombinant yeast HUB1 was tested by immunoblot against yeast lysates expressing the HUB1-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 Hub1 using a 1:500 dilution of HUB1 polyclonal antibody (Cat # PAB11361) (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.
Yeast lysate proteins were separated by SDS-PAGE using a 15% gel. This data indicates that anti-HUB1 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.

  • Specifications

    Product Description

    Rabbit polyclonal antibody raised against recombinant HUB1.

    Immunogen

    Recombinant protein corresponding to Saccharomyces cerevisiae HUB1.

    Host

    Rabbit

    Reactivity

    Yeast

    Form

    Lyophilized

    Quality Control Testing

    Antibody Reactive Against Recombinant Protein.

    Recommend Usage

    ELISA (1:1000-1:5000)
    Western Blot (1:500)
    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 HUB1 fusion protein.
    HUB1 polyclonal antibody (Cat # PAB11361) generated by immunization with recombinant yeast HUB1 was tested by immunoblot against yeast lysates expressing the HUB1-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 Hub1 using a 1:500 dilution of HUB1 polyclonal antibody (Cat # PAB11361) (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.
    Yeast lysate proteins were separated by SDS-PAGE using a 15% gel. This data indicates that anti-HUB1 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

  • Gene Info — HUB1

    Entrez GeneID

    855767

    Protein Accession#

    Q6Q546

    Gene Name

    HUB1

    Gene Alias

    -

    Gene Description

    Hub1p

    Gene Ontology

    Hyperlink

    Gene Summary

    may function in modification of Sph1p and Hbt1p

    Other Designations

    Ubiquitin-like protein modifier, may function in modification of Sph1p and Hbt1p, functionally complemented by the human or S. pombe ortholog; mechanism of Hub1p adduct formation not yet clear

  • Publication Reference