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Tags & Cell Markers Fusion / Marker Proteins GAL4

Anti-GAL4抗体(ab1396)

  • Datasheet
  • SDS
Reviews (2)Q&A (2)References (10)

Product price, shipping and contact information

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Western blot - Anti-GAL4 antibody (ab1396)
  • ChIP - Anti-GAL4 antibody (ab1396)
  • ChIP - Anti-GAL4 antibody (ab1396)
  • Western blot - Anti-GAL4 antibody (ab1396)

Key features and details

  • Rabbit polyclonal to GAL4
  • Suitable for: WB, ChIP
  • Reacts with: Saccharomyces cerevisiae
  • Isotype: IgG

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概述

  • 产品名称

    Anti-GAL4抗体
    参阅全部 GAL4 一抗
  • 描述

    兔多克隆抗体to GAL4
  • 宿主

    Rabbit
  • 特异性

    Customers feedbacks suggests that this antibody would not provide satisfactory results in Drosophila melanogaster.
  • 经测试应用

    适用于: WB, ChIPmore details
  • 种属反应性

    与反应: Saccharomyces cerevisiae
    不与反应: Drosophila melanogaster
  • 免疫原

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

  • 阳性对照

    • This antibody gave a positive signal with GAL4-VP16 recombinant protein.
  • 常规说明

    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 the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

性能

  • 形式

    Liquid
  • 存放说明

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • 存储溶液

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • 纯度

    Immunogen affinity purified
  • 克隆

    多克隆
  • 同种型

    IgG
  • 研究领域

    • Tags & Cell Markers
    • Fusion / Marker Proteins
    • GAL4
    • Signal Transduction
    • Metabolism
    • Energy Metabolism
    • Signal Transduction
    • Cytoskeleton / ECM
    • Cell Adhesion
    • Gap Junctions
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Polymerase associated factors
    • Pol II Transcription
    • Other
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Other factors
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Transcription Factors
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Carbohydrate metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Energy transfer pathways
    • Energy Metabolism
    • Metabolism
    • Types of disease
    • Cancer

相关产品

  • ChIP Related Products

    • Rabbit Anti-Mouse IgG H&L (ab46540)
    • ChIP Kit (ab500)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant Human GAL4 protein (ab89234)

应用

The Abpromise guarantee

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.

应用 Ab评论 说明
WB
Use a concentration of 1 µg/ml. Predicted molecular weight: 99 kDa.
ChIP
Use at an assay dependent concentration.
说明
WB
Use a concentration of 1 µg/ml. Predicted molecular weight: 99 kDa.
ChIP
Use at an assay dependent concentration.

靶标

  • 相关性

    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.
  • 细胞定位

    Nuclear
  • 数据库链接

    • Entrez Gene: 855828 Saccharomyces cerevisiae
    • SwissProt: P04386 Saccharomyces cerevisiae
    • 别名

      • Gal4p antibody
      • GAL81 antibody
      • Regulatory protein GAL4 antibody

    图片

    • Western blot - Anti-GAL4 antibody (ab1396)
      Western blot - Anti-GAL4 antibody (ab1396)

      Anti-GAL4 (ab1396) antibody at 1ug/ml.

      Lane 1: Marker.

      Lane 2A: Whole cell yeast lysate control from untransformed SEY6210 pep4-3 strain (a kind gift from Prof. Tom Stevens from University of Oregon).

      Lane 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).

      Lane 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).

      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)

      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.

    • ChIP - Anti-GAL4 antibody (ab1396)
      ChIP - Anti-GAL4 antibody (ab1396)This image is courtesy of the Tony Kouzarides lab at the Gurdon Institute.
      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).
    • ChIP - Anti-GAL4 antibody (ab1396)
      ChIP - Anti-GAL4 antibody (ab1396)This image is courtesy of an anonymous collaborator.

      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.

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

      Secondary
      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.

    实验方案

    • ChIP protocols
    • Western blot protocols

    Click here to view the general protocols

    数据表及文件

    • SDS download

    • Datasheet download

      Download

    文献 (10)

    发表研究结果有使用 ab1396?请让我们知道,以便我们可以引用本数据表中的参考文章。

    ab1396 被引用在 10 文献中.

    • Elison GL  et al. Insights into Bidirectional Gene Expression Control Using the Canonical GAL1/GAL10 Promoter. Cell Rep 25:737-748.e4 (2018). 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 . 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 . PubMed: 26173127
    • Yang J  et al. GEMIN4 functions as a coregulator of the mineralocorticoid receptor. J Mol Endocrinol 54:149-60 (2015). PubMed: 25555524
    • Chen X  et al. Activation of a human chromosomal replication origin by protein tethering. Nucleic Acids Res 41:6460-74 (2013). WB . PubMed: 23658226
    • Inturi R  et al. Adenovirus precursor pVII protein stability is regulated by its propeptide sequence. PLoS One 8:e80617 (2013). WB . PubMed: 24260437
    • Dawson MA  et al. Three distinct patterns of histone H3Y41 phosphorylation mark active genes. Cell Rep 2:470-7 (2012). WB . PubMed: 22999934
    • Knutson BA & Hahn S Domains of Tra1 important for activator recruitment and transcription coactivator functions of SAGA and NuA4 complexes. Mol Cell Biol 31:818-31 (2011). ChIP ; Saccharomyces cerevisiae . PubMed: 21149579
    • Pinskaya M  et al. H3 lysine 4 di- and tri-methylation deposited by cryptic transcription attenuates promoter activation. EMBO J 28:1697-707 (2009). ChIP ; Saccharomyces cerevisiae . PubMed: 19407817
    • Ghosh M  et al. Transcription factor binding and induced transcription alter chromosomal c-myc replicator activity. Mol Cell Biol 24:10193-207 (2004). WB . PubMed: 15542830

    客户评价及客户问答

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    1-4 of 4 Abreviews or Q&A

    IHC - Wholemount abreview for Anti-GAL4 antibody - ChIP Grade

    Poor
    Abreviews
    Abreviews
    This product is known to not work in this application or species.
    abreview image
    Application
    IHC - Wholemount
    Sample
    Fruit fly (Drosophila melanogaster) Tissue (Wholemount brain tissue)
    Specification
    Wholemount brain tissue
    Read More

    Abcam user community

    Verified customer

    提交于 Jan 06 2014

    Question

    Inquiry: The customer is looking for the positive control for ab1396 for ChIP. Is it availavble for the purchase from Abcam? She prefers the lysate of the cells that express Gal4 phused protein. Are there any other alternatives? Thank you for the assistance. Have a wonderful day.

    Read More

    Abcam community

    Verified customer

    Asked on Jun 20 2012

    Answer

    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!
    https://www.abcam.com/abreviews

    Read More

    Abcam Scientific Support

    回复于 Jun 20 2012

    Immunohistochemistry (Frozen sections) abreview for Anti-GAL4 antibody - ChIP Grade

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

    Ms. Vilaiwan Fernandes

    Verified customer

    提交于 Nov 16 2011

    Question

    that ain't no fly gene from the Expasy blast, it's the yeast gene (unless I'm profoundly mistaken...). The sole reference in the Expasy output is to a paper entitled "Dual-tagging gene trap of novel genes in Drosophila melanogaster";.meaning the hit was a transgene derived from the yeast Gal4 gene. This is not surprising inasmuch as Gal4 is extensively used in the fly to express UAS-driven transgenes... Thanks for looking into things for me.

    Read More

    Abcam community

    Verified customer

    Asked on Feb 25 2005

    Answer

    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.

    Read More

    Abcam Scientific Support

    回复于 Mar 02 2005

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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