• Product name
    Anti-GAL4 antibody - ChIP Grade
    See all GAL4 primary antibodies
  • Description
    Rabbit polyclonal to GAL4 - ChIP Grade
  • Host species
  • Specificity
    Customers feedbacks suggests that this antibody would not provide satisfactory results in Drosophila melanogaster.
  • Tested applications
    Suitable for: WB, ChIPmore details
  • Species reactivity
    Reacts with: Saccharomyces cerevisiae
    Does not react with: Drosophila melanogaster
  • Immunogen

    Synthetic peptide corresponding to Saccharomyces cerevisiae GAL4 aa 100-200 conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab23612)

  • Positive control
    • This antibody gave a positive signal with GAL4-VP16 recombinant protein.


Our Abpromise guarantee covers the use of ab1396 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.
ChIP Use at an assay dependent dilution.


  • 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
  • Database links
    • Alternative names
      • Gal4p antibody
      • GAL81 antibody
      • Regulatory protein GAL4 antibody


    • Developed using the ECL technique. Performed under reducing conditions with exposure time of 5 mins. The samples run on a 4-20% gradient gel. All blocking and antibody incubation steps done in 5% milk in 20mM Tris-HCl plus 0.1% TWEEN-20.
      Sample 1: Marker. Sample 2A: Whole cell yeast lysate control from untransformed SEY6210 pep4-3 strain (a kind gift from Prof. Tom Stevens from University of Oregon). Sample 2B: 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 from University of Oregon). Sample 2C: 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 from University of Oregon).
      Primary: Lane 1: none. Lane 2: Anti-GAL4BD (ab1396) antibody at 1ug/ml. Secondary: HRP conjugated Goat anti-Rabbit secondary antibody at 1/10000 dilution. Predicted band size: 99 kDa. Additional bands: 56 kDa (possible isoform, Fusion Protein)
    • A stably transfected 293T human cell line harbouring the GAL4 upstream activation sequence was transiently transfected with a V5 or T7- tagged GAL4 DNA Binding Domain construct. 48 hours post transfection Chromatin was prepared according to the Abcam X-ChIP protocol. The ChIP was performed with 25 ug chromatin, 5ug of antibody and 20 ul of Protein A/G beads. A non-specific antibody was used as the negative control. The immunoprecipitated DNA was quantified by real time PCR (SYBR Green approach).
    • S. cerevisiae cells were incubated in raffinose-containing media then transferred to galactose-containing media to activate transcription of galactose activated genes. IP was performed with 500µl of cell extract incubated overnight with 5µl of ab1396 at 4°C, followed by the addition of 25µl of protein A sepharose beads and incubation for 2 hours (room temp). The subsequently purified DNA was analysed with 3 pairs of primers (see diagram). The results show that Gal4 binds specifically to the UAS region of Gal1-Gal10 in both raffinose and galactose, despite only 500-600bp between the UAS and 5' primer.

    • Anti-GAL4 antibody - ChIP Grade (ab1396) at 1 µg/ml + GAL4-VP16 Recombinant Protein at 0.1 µg

      Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

      Performed under reducing conditions.

      Predicted band size: 99 kDa
      Observed band size: 32 kDa
      why is the actual band size different from the predicted?

      Exposure time: 2 minutes

      ab1396 was tested against GAL4-VP16 Recombinant Protein predicted to run at 28 kDa.


    This product has been referenced in:
    • Elison GL  et al. Insights into Bidirectional Gene Expression Control Using the Canonical GAL1/GAL10 Promoter. Cell Rep 25:737-748.e4 (2018). Read more (PubMed: 30332652) »
    • Li J  et al. Development of a membrane-anchored ligand and receptor yeast two-hybrid system for ligand-receptor interaction identification. Sci Rep 6:35631 (2016). WB . Read more (PubMed: 27762338) »
    See all 10 Publications for this product

    Customer reviews and Q&As

    1-4 of 4 Abreviews or Q&A

    This product is known to not work in this application or species.
    IHC - Wholemount
    Fruit fly (Drosophila melanogaster) Tissue (Wholemount brain tissue)
    Wholemount brain tissue

    Abcam user community

    Verified customer

    Submitted Jan 06 2014


    Thank you for contacting us.
    We have 2 Gal4 products available in catalogue. One is ab23612 which was used to produce ab1396 and other is ab81879 which is a fusion protein. These products can be used a positive control.
    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.
    Use our products? Submit an Abreview. Earn rewards!

    Read More
    This product is known to not work in this application or species.
    Immunohistochemistry (Frozen sections)
    Fruit fly (Drosophila melanogaster) Tissue sections (eye imaginal disc)
    eye imaginal disc
    Yes - tritonX
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: ~20°C

    Ms. Vilaiwan Fernandes

    Verified customer

    Submitted Nov 16 2011

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