Overview

  • Product name

    Anti-GAL4 antibody [15-6E10A7]
    See all GAL4 primary antibodies
  • Description

    Mouse monoclonal [15-6E10A7] to GAL4
  • Host species

    Mouse
  • Tested applications

    Suitable for: WB, Othermore details
  • Species reactivity

    Reacts with: Saccharomyces cerevisiae
  • Immunogen

    Recombinant fragment. This information is considered to be commercially sensitive.

  • General notes

    This antibody clone is manufactured by Abcam.

    The GAL4 DNA-BD monoclonal antibody binds specifically to the DNA-binding domain of the yeast GAL4 protein. This antibody is useful for verifying the expression and correct molecular weight of bait fusion proteins containing GAL4 DNA BD as the fusion partner in yeast two hybrid studies. The binding domain is typically used in a two-hybrid experiment as the fusion partner for the protein of interest. When the fusion protein of interest cannot activate transcription on its own, it can be used as bait to screen a library of cDNA clones that are fused to an activation domain. GAL4 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.

    Product was previously marketed under the MitoSciences sub-brand.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C.
  • Storage buffer

    Preservative: 0.02% Sodium azide
  • Concentration information loading...
  • Purity

    Ammonium Sulphate Precipitation
  • Primary antibody notes

    The GAL4 DNA-BD monoclonal antibody binds specifically to the DNA-binding domain of the yeast GAL4 protein. This antibody is useful for verifying the expression and correct molecular weight of bait fusion proteins containing GAL4 DNA BD as the fusion partner in yeast two hybrid studies. The binding domain is typically used in a two-hybrid experiment as the fusion partner for the protein of interest. When the fusion protein of interest cannot activate transcription on its own, it can be used as bait to screen a library of cDNA clones that are fused to an activation domain. GAL4 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.
  • Clonality

    Monoclonal
  • Clone number

    15-6E10A7
  • Isotype

    IgG1
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab135397 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
WB Use a concentration of 1 µg/ml. Predicted molecular weight: 99 kDa.
Other Use at an assay dependent concentration. Yeast two hybrid studies

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

    Images

    • All lanes : Anti-GAL4 antibody [15-6E10A7] (ab135397) at 1 µg/ml

      Lane 1 : Whole cell yeast lysate control from untransformed SEY6210 pep4-3 strain (a kind gift from Prof. Tom Stevens, University of Oregon).
      Lane 2 : Whole cell yeast lysate from SEY6210 pep4-3 strain transformed with pJK22-pGBDU plasmid containing GAL4 DNA Binding domain fused to the VPS60 Gene (a kind gift from Prof. Tom Stevens, University of Oregon).
      Lane 3 : Whole cell yeast lysate from SEY6210 pep4-3 strain transformed with pJk23-pGAD plasmid containing GAL4 Active Domain fused to the VPS60 Gene (a kind gift from Prof. Tom Stevens, University of Oregon).

      Secondary
      All lanes : Anti-Mouse IgG (H&L) Goat Polyclonal

      Performed under reducing conditions.

      Predicted band size: 99 kDa
      Additional bands at: 56 kDa (possible isoform), 56 kDa (possible Fusion protein)


      Exposure time: 3 minutes

    References

    This product has been referenced in:

    • Rother S  et al. NF-?B-repressing factor phosphorylation regulates transcription elongation via its interactions with 5'?3' exoribonuclease 2 and negative elongation factor. FASEB J 30:174-85 (2016). WB . Read more (PubMed: 26340924) »
    • Geng S  et al. Protein Interaction between Ameloblastin and Proteasome Subunit a Type 3 Can Facilitate Redistribution of Ameloblastin Domains within Forming Enamel. J Biol Chem 290:20661-73 (2015). Read more (PubMed: 26070558) »
    See all 2 Publications for this product

    Customer reviews and Q&As

    1-2 of 2 Abreviews or Q&A

    Answer

    Thank you for your email.

    We scientific support team at Abcam have years of experience in laboratory as part of our PhDs and research projects. We do not make fun of our customers as we know how it feels when research gets stuck just due to a faulty or wrong product so we try to be very lenient with customers in order to help them, in their research.

    As of ab1396 datasheet we list predicted species on datasheets if the proteins show 80-90% homology. Although the GAL4 protein is not expressed in Drosophila however because similar UAS-GAL4 expression system is present in drosophila and if the same S.cereveisiae fragment is expressed then why the antibody shouldn't cross react. This was the idea of keeping Drosophila as predicted species, which I have removed now.

    I am happy to replace this antibody with ab9110. Could you send me the purchase order number of ab1396?

    Many thanks for your cooperation in this case. I will look forward to hearing from you soon.

    Read More

    Answer

    Thank you very much for your patience.

    I had requested a retest of this antibody with the wild type and transfected cell lysates we have. I can confirm that the antibody (ab1396, ab135397 and ab135398) is specific to the target and does not bind to control wild type cell line (no insert). The lysates are same as of ab135398.

    Please find attached the results.

    I hope these results will helpful to retest the wild type cell line you are using. Please do not hesitate to contact me with any further question.

    Read More

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