Overview

  • Product name
    Anti-GAL4 antibody [GAL4–8]
    See all GAL4 primary antibodies
  • Description
    Mouse monoclonal [GAL4–8] to GAL4
  • Host species
    Mouse
  • Specificity
    This antibody can be used to detect both N-terminal and C-terminal tagged Gal4(1–147) and Gal4(1–92) fusion proteins. The clone number has been updated from (3G152) to (GAL4–8) both clone numbers name the same antibody clone.
  • Tested applications
    Suitable for: ICC/IF, ELISA, WB, ICCmore details
  • Species reactivity
    Reacts with: Saccharomyces cerevisiae
  • Immunogen

    Recombinant fragment, corresponding to amino acids 1-147 of S. cerevisiae GAL4.

  • Epitope
    The immunoreactive epitope maps within amino acids 1–92 of GAL4.
  • Positive control
    • N-terminal and C-terminal tagged GAL4 (1-147) and GAL4 (1-92) fusion proteins

Properties

Applications

Our Abpromise guarantee covers the use of ab14477 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
ICC/IF Use at an assay dependent dilution. PubMed: 20610754
ELISA Use a concentration of 0.1 - 1 µg/ml.
WB Use a concentration of 3 µg/ml. Predicted molecular weight: 99 kDa.
ICC Use at an assay dependent concentration.

Target

  • Relevance
    Function: This protein is a positive regulator for the gene expression of the galactose-induced genes such as GAL1, GAL2, GAL7, GAL10, and MEL1 which encode for the enzymes used to convert galactose to glucose. It recognizes a 17 base pair sequence in (5'-CGGRNNRCYNYNCNCCG-3') the upstream activating sequence (UAS-G) of these genes. Subunit structure: Binds DNA as a homodimer. Interacts directly with the mediator subunits GAL11/MED15 and SRB4/MED17. Domain: The 9aaTAD motif (residues 862 to 870) is a transactivation domain present in a large number of yeast and animal transcription factors. Post-translational modification: Association between GAL11 and GAL4 may serve to expedite phosphorylation of GAL4.
  • Cellular localization
    Nuclear
  • Database links
    • Alternative names
      • Gal4p antibody
      • GAL81 antibody
      • Regulatory protein GAL4 antibody

    References

    This product has been referenced in:
    • Ren R  et al. Visualization of aging-associated chromatin alterations with an engineered TALE system. Cell Res 27:483-504 (2017). Read more (PubMed: 28139645) »
    • Chen Y  et al. Intramembranous valine linked to schizophrenia is required for neuregulin 1 regulation of the morphological development of cortical neurons. J Neurosci 30:9199-208 (2010). ICC/IF . Read more (PubMed: 20610754) »
    See all 2 Publications for this product

    Customer reviews and Q&As

    1-4 of 4 Abreviews or Q&A

    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    IHC - Wholemount
    Sample
    Drosophila melanogaster Tissue (Imaginal Wing Disc)
    Specification
    Imaginal Wing Disc

    Mr. Michael Baumgartner

    Verified customer

    Submitted Apr 17 2018

    Answer

    Thisantibody has only been tested with GAL4-fusion proteins inWestern Blot.

    I hope this is helpful. Please contact me again if you have any further questions.

    Read More
    Application
    Immunocytochemistry
    Sample
    Human Recombinant protein (Transfected cells)
    Specification
    Transfected cells
    Fixative
    Paraformaldehyde
    Blocking step
    Serum as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: 5%

    Abcam user community

    Verified customer

    Submitted Aug 17 2005

    Answer

    Thank you for contacting us for technical support and taking the time to fill in our protocol questionnaire, I'm very sorry to hear you are experiencing problems with ab14477 in your samples. I have looked at your order details and can confirm there were no shipping delays suggesting the antibody is damaged, we have not received any other complaints about this antibody. I would like to recommend the following modifications to your protocol to maximise the signal: - load more protein per well (e.g 40ug) -incubate the antibody at a higher concentration, the 3ug/ml concentration is a recommendation and you may need to try 5ug/ml. -the dilution/blocking buffer you mention might be the problem as it has not been tested with this antibody. I would definitely try two blocking agents: try 5%BSA (1hr) and 5%milk (1hr) and dilute the antibodies in TBST (Tween 0.1%v/v in Tris buffer) -you do not mention a lysis buffer, does it contain fresh protease inhibitors? Finally I am not familiar with your secondary antibody but it may need to be a bit more concentrated (e.g 1:3000-1:5000)? I hope these suggestions will help you get a better signal, if you still experience problems please do not hesitate to contact me again,

    Read More

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