• 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:
    • 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) »
    • Roedgaard M  et al. DNA Topoisomerases Are Required for Preinitiation Complex Assembly during GAL Gene Activation. PLoS One 10:e0132739 (2015). WB, ChIP ; Saccharomyces cerevisiae . Read more (PubMed: 26173127) »

    See all 9 Publications for this product

    Customer reviews and Q&As

    Abcam has not validated the combination of species/application used in this Abreview.
    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...

    Read More
    Abcam has not validated the combination of species/application used in this Abreview.
    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

    Thanks for your email. Yes, I do think you're right. I removed Drosophila from the datasheets. If you have any other questions or concerns, just let me know.


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