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Anti-GAL4 antibody [14-7E10G10] (ab135398)

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Western blot - Anti-GAL4 antibody [14-7E10G10] (ab135398)

    Key features and details

    • Mouse monoclonal [14-7E10G10] to GAL4
    • Suitable for: WB
    • Reacts with: Saccharomyces cerevisiae
    • Isotype: IgG2b

    Overview

    • Product name

      Anti-GAL4 antibody [14-7E10G10]
      See all GAL4 primary antibodies

    • Description

      Mouse monoclonal [14-7E10G10] to GAL4

    • Host species

      Mouse

    • Tested applications

      Suitable for: WBmore details

    • Species reactivity

      Reacts with: Saccharomyces cerevisiae

    • Immunogen

      Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

    • General notes

      The GAL4 Activation Domain monoclonal antibody binds specifically to the AD of the yeast GAL4 protein. This antibody is useful for verifying the expression and correct molecular weight of fusion proteins containing GAL4 AD as the fusion partner in two hybrid studies. The activation domain is typically used in a two-hybrid experiment as the fusion partner for a library of cDNA clones to screen against a protein of interest fused to a DNA-binding domain. Gal4is 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.

      This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

      This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

      The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

      If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

    Properties

    • Form

      Liquid

    • Storage instructions

      Shipped at 4°C. Store at +4°C.

    • Storage buffer

      pH: 7.5
      Preservative: 0.02% Sodium azide
    • Concentration information loading...

    • Purity

      Ammonium Sulphate Precipitation

    • Primary antibody notes

      The GAL4 Activation Domain monoclonal antibody binds specifically to the AD of the yeast GAL4 protein. This antibody is useful for verifying the expression and correct molecular weight of fusion proteins containing GAL4 AD as the fusion partner in two hybrid studies. The activation domain is typically used in a two-hybrid experiment as the fusion partner for a library of cDNA clones to screen against a protein of interest fused to a DNA-binding domain. Gal4is 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.

    • Clonality

      Monoclonal

    • Clone number

      14-7E10G10

    • Isotype

      IgG2b

    • Research areas

    Applications

    The Abpromise guarantee

    Our Abpromise guarantee covers the use of ab135398 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 0.5 µg/ml. Predicted molecular weight: 99 kDa.
    Notes
    WB
    Use a concentration of 0.5 µg/ml. Predicted molecular weight: 99 kDa.

    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

      • Western blot - Anti-GAL4 antibody [14-7E10G10] (ab135398)
        Western blot - Anti-GAL4 antibody [14-7E10G10] (ab135398)
        All lanes : Anti-GAL4 antibody [14-7E10G10] (ab135398) at 0.5 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
        Lane 1 : Goat polyclonal to Mouse IgG – H&L – Pre-Adsorbed (HRP) - 1:10000
        Lane 2 : Goat polyclonal to Mouse IgG – H&L – Pre-Adsorbed (HRP)- 1:10000
        Lane 3 : Goat polyclonal to Mouse IgG – H&L – Pre-Adsorbed (HRP) - 1:10000

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

      References (0)

      Publishing research using ab135398? Please let us know so that we can cite the reference in this datasheet.

      ab135398 has not yet been referenced specifically in any publications.

      Customer reviews and Q&As

      Show All Reviews Q&A
      Submit a review Submit a question

      1-2 of 2 Abreviews or Q&A

      Question

      Ok, at last we do understand each other. I am sorry for the harsh tone
      in my last e-mail, I just had the impression you want to make fun of
      me showing me those unrelated test results.Please, next time just say
      that you don't have data and/or that you will not further investigate
      into the matter. This would have been a fair answer.
      We still do not know which Antibody to use. This is the 3rd we tried.
      SC-577 (works in epifluorescence but stains unspecifically on
      Westerns), ab3G152 (works neither in epifluorescence nor on Westerns)
      and ab1396 (only tested on Westerns with the results mentioned). For
      sure we will not waste time with a fourth one and detect Gal4 by Mass
      Spec directly.
      In a previous Mail you wrote "I am happy to replace the lot with a
      different one or a different antibody (please choose from catalogue)
      and finally can refund the money you have paid for ab1936". We use
      ab9110 which performs well. A discount for this antibody would help us
      most.
      Thank you for your efforts.

      Read More
      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

      Question

      Product code: 1396
      Lot number: GR46134-1
      Inquiry: We used ab1396 on Drosophila cell culture and embryonic lysates. We do get a band at around 35 kDa, unfortunately also in the negative control of cells/flies that do not harbour the yeast Gal4 transgene. Western Blot is attached.

      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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