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R718 Mouse Anti-GATA3
R718 Mouse Anti-GATA3
Flow cytometric analysis of GATA3 expression in Jurkat cells. Cells from the human Ramos (Burkitt's lymphoma, ATCC CRL-1596; dashed line histogram) and Jurkat (Acute T cell leukemia, ATCC TIB-152; solid line histogram) cell lines were fixed (10 min, 37ºC) with pre-warmed BD Cytofix™ Buffer (Cat. No. 554655) and then permeabilized (on ice, 30 min) with BD Phosflow™ Perm Buffer III (Cat. No. 558050). The cells were washed with BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656) and stained with BD Horizon™ R718 Mouse Anti-GATA3 antibody (Cat. No. 567632/567633). The fluorescence histograms showing GATA3 expression were derived from gated events with the forward and side light-scatter characteristics of intact lymphoid cells. Flow cytometric analysis was performed using a LSRFortessa™ X-20 Flow Cytometer System and FlowJo™ software.
Flow cytometric analysis of GATA3 expression in Jurkat cells. Cells from the human Ramos (Burkitt's lymphoma, ATCC CRL-1596; dashed line histogram) and Jurkat (Acute T cell leukemia, ATCC TIB-152; solid line histogram) cell lines were fixed (10 min, 37ºC) with pre-warmed BD Cytofix™ Buffer (Cat. No. 554655) and then permeabilized (on ice, 30 min) with BD Phosflow™ Perm Buffer III (Cat. No. 558050). The cells were washed with BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656) and stained with BD Horizon™ R718 Mouse Anti-GATA3 antibody (Cat. No. 567632/567633). The fluorescence histograms showing GATA3 expression were derived from gated events with the forward and side light-scatter characteristics of intact lymphoid cells. Flow cytometric analysis was performed using a LSRFortessa™ X-20 Flow Cytometer System and FlowJo™ software.
Product Details
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BD Horizon™
Gata-3; GATA binding protein 3; GATA-binding factor 3; HDR
Human (QC Testing), Mouse (Tested in Development)
Mouse BALB/c IgG1, κ
Conserved peptide between the trans-activation and DNA-binding domains of human, mouse and rat GATA3
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unreacted dye was removed.

Recommended Assay Procedures

  BD® CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

Product Notices

  1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  4. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  5. This product is provided under an Agreement between BIOTIUM and BD Biosciences. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
  6. Alexa Fluor™ is a trademark of Life Technologies Corporation.
  7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  8. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
567633 Rev. 1
Antibody Details
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L50-823

GATA3 (GATA binding protein 3) is a member of the GATA family of transcription factors. This ~50-kDa nuclear protein regulates the development and subsequent maintenance of multiple tissues. GATA3 is involved in the development of T lymphocytes (regulates T cell receptor subunit gene expression) and the differentiation of mature T cells to become Th2 cells. The expressed levels of normal or mutant GATA3 are also associated with the behaviors of various cancer cells including estrogen receptor-positive breast carcinoma cells.

The L50-823 monoclonal antibody recognizes human and mouse GATA3.

  

The antibody was conjugated to BD Horizon™ Red 718, which has been developed exclusively by for BD Biosciences as a better alternative to Alexa Fluor™ 700. BD Horizon™ Red 718 can be excited by the red laser (628 – 640 nm) and, with an Em Max around 718 nm, it can be detected using a 730/45 nm filter. Due to similar excitation and emission properties, we do not recommend using R718 in combination with APC-R700 or Alexa Fluor™ 700.

567633 Rev. 1
Format Details
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R718
The BD Horizon™ Red 718 (R718) Dye is part of the BD red family of dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 695-nm and an emission maximum (Em Max) at 718-nm. Driven by BD innovation, R718 is designed to be excited by the red laser (627–640-nm) and detected using an optical filter centered near 720-nm (e.g., a 720/40-nm bandpass filter). R718 is a brighter alternative to Alexa Fluor™ 700. R718 is also a bright small molecule alternative to APC-R700 with lower spread into the APC detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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R718
Red 627-640 nm
695 nm
718 nm
567633 Rev.1
Citations & References
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Development References (11)

  1. Asselin-Labat M-L, Sutherland KD, Barker H, et al. Gata-3 is an essential regulator of mammary-gland morphogenesis and luminal-cell differentiation. Nat Cell Biol. 2006; 9:201-209. (Biology).
  2. Delgoffe GM, Pollizzi KN, Waickman AT, et al. The kinase mTOR regulates the differentiation of helper T cells through the selective activation of signaling by mTORC1 and mTORC2. Nat Immunol. 2011; 12(4):295-303. (Clone-specific: Flow cytometry). View Reference
  3. Kouros-Mehr H, Slorach EM, Sternlicht MD, Werb Z. GATA-3 maintains the differentiation of the luminal cell fate in the mammary gland. Cell. 2006; 127:1041-1055. (Biology).
  4. Lesourne R, Uehara S, Lee J, et al. Themis, a T cell-specific protein important for late thymocyte development. Nat Immunol. 2009; 10(8):840-847. (Clone-specific: Flow cytometry). View Reference
  5. Marine J, Winoto A. The human enhancer-binding protein Gata3 binds to several T-cell receptor regulatory elements. Proc Natl Acad Sci U S A. 1991; 88(16):7284-7288. (Biology).
  6. Miyamoto H, Izumi K, Yao JL, et al. GATA binding protein 3 is down-regulated in bladder cancer yet strong expression is an independent predictor of poor prognosis in invasive tumor. Hum Pathol. 2012; 43(11):2033-2040. (Clone-specific: Immunohistochemistry). View Reference
  7. Steenbergen RDM, OudeEngberink VE, Kramer D, et al. Down-regulation of GATA-3 expression during human papillomavirus-mediated immortalization and cervical carcinogenesis. Am J Pathol. 2002; 160(6):1945-1951. (Biology). View Reference
  8. Usary J, Llaca V, Karaca G, et al. Mutation of GATA3 in human breast tumors. Oncogene. 2004; 23(46):7669-7678. (Biology). View Reference
  9. Yang Z, Gu L, Romeo P-H, et al. Human GATA-3 trans-activation, DNA-binding, and nuclear localization activities are organized into distinct structural domains. Mol Cell Biol. 1994; 14(3):2201-2212. (Biology). View Reference
  10. Zheng W, Flavell RA. The transcription factor GATA-3 is necessary and sufficient for Th2 cytokine gene expression in CD4 T cells. Cell. 1997; 89(4):587-596. (Biology). View Reference
  11. van Esch H, Groenen P, Nesbit MA, et al. GATA3 haplo-insufficiency causes human HDR syndrome. Nature. 2000; 106:419-422. (Biology). View Reference
View All (11) View Less
567633 Rev. 1

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.