GATA3 Antibody | 1A12-1D9

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GATA3 Antibody | 1A12-1D9 gallery image 1

Western blot analysis of GATA3 was performed by loading 15ug of SH-SY5Y whole cell lysate (WCL) and SH-SY5Y nuclear extract (NE) onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked for at least 1 hour. The membrane was probed with Mouse anti GATA3 (MCA6036) at a dilution of 1/1000 overnight at 4°C on a rocking platform, washed in TBS-0.1%Tween-20, and probed with an HRP-conjugated goat anti-mouse IgG secondary antibody for 1 hour. Chemiluminescent detection was performed

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Immunofluorescent analysis of GATA3 (green) in MCF7 cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature. Cells were blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed without (left panel) or with (right panel) Mouse anti GATA3 (MCA6036) at a dilution of 1/50 for at least 1 hour at room temperature, washed with PBS, and incubated with a DyLight 488-conjugated goat anti-mouse IgG secondary antibody for 30 minutes at room temperature. F-Actin (red) was stained with phalloidin (red) and nuclei (blue) were stained with Hoechst 33342 dye. Images were taken at 20x magnification

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Immunohistochemistry analysis of GATA3 showing staining in the nucleus of paraffin-embedded human breast carcinoma (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with dH2O and PBS, and then probed with a Mouse anti GATA3 (MCA6036) diluted in 3% BSA-PBS at a dilution of 1/1000 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting

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  • Mouse anti GATA3
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  • Product Type
    Monoclonal Antibody
  • Clone
    1A12-1D9
  • Isotype
    IgG1
1 Formats Available
    Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
    MCA6036IC, IF, IP, P *, WBdatasheet pdfdatasheet pdf50 µg
    MCA6036
    Summary
    Secondary Antibodies
    Negative Isotype Controls
    Useful Reagents
    Positive Controls
    Histology Controls
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    References
    Reviews
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    • Mouse anti GATA3 antibody clone 1A12-1D9 recognizes GATA3 also known as trans-acting T-cell-specific transcription factor GATA-3 or GATA-binding factor 3.

      The GATA3 protein is a member of the GATA family of transcription factors which has received its name from binding DNA sequences containing the GATA consensus motif 5'-[AT]GATA[AG]-3' (P15976). GATA3 has been shown to play a critical role in T cell development and the differentiation of T helper 2 (Th2) cells (Ho et al. 1991). In addition, GATA3 has been implied to have tissue specific functions such as a regulatory role in the development of the mammary gland epithelium and skin tissue (Chou et al. 2010).

      Aberrant GATA3 levels have also been identified in a variety of cancers including breast cancer. Indeed, GATA3 has become a marker for certain breast cancer types and GATA3 antibodies are commonly used for the immunohistochemical evaluation of breast cancer samples (Krings et al. 2014).
    • This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.

    • Application NameYesNoMin DilutionMax Dilution
      Immunocytochemistry1/501/100
      Immunofluorescence1/501/100
      Immunoprecipitation5 ug
      Western Blotting1/1000
      Immunohistology - Paraffin(1)1/2001/2000
      (1)
      This antibody requires heat-mediated antigen retrieval prior to staining. Citrate buffer pH6.0 is recommended for this purpose.

    • Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
    • Technical Advice
    • Recommended Protocol
    • ELISA
    • Immunohistology
    • Histology Positive Control Tissue
    • Immunofluorescence
    • Western Blotting
      Mouse anti GATA3 antibody recognizes a band of approximately 55 kDa in SH-SY5Y cell lysate and SH-SY5Y nuclear extract
    • Instructions For Use

    Additional GATA3 Antibody Formats

    Formats Clone Applications Sizes available
    GATA3 Antibody : Purified 1A12-1D9 IC, IF, IP, P *, WB 50 µg
    • Copyright © 2016 Bio-Rad

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      Recommended Negative Isotype Control

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