Comparison of GATA3 expression in human T and B cell lines. Jurkat T leukemia (ATCC TIB152, shaded histogram) and Ramos Burkitt's lymphoma (ATCC CRL-1596, open histogram) were fixed with pre-warmed BD Cytofix™ buffer (Cat. No. 554655) for 10 minutes at 37ºC, permeabilized with BD™ Phosflow Perm Buffer III (Cat. No. 558050) on ice for at least 30 minutes, and then stained with either PE Mouse anti-GATA3 or PE Mouse IgG1 κ Isotype control (Cat. No. 559320, not shown). The GATA3 staining on the Jurkat cell line was significantly brighter than the isotype control on Jurkat cells, while the GATA3 staining on the Ramos cells coincided very closely to its isotype control (data not shown). Thus, GATA3 expression was detected on the T cell line but not the B cell line. Flow cytometry was performed on a BD FACSArray™ bioanalyzer system.
Comparison of GATA3 expression in mouse Th2 and Th1 cell lines. D10.G4.1 Th2 lymphoblasts (ATCC TIB-224, shaded histogram) and 2D6 Th1 clone (Ahn et al, 1998, open histogram) were fixed with pre-warmed BD Cytofix™ buffer (Cat. No. 554655) for 10 minutes at 37ºC, permeabilized with BD™ Phosflow Perm Buffer III (Cat. No. 558050) on ice for at least 30 minutes, and then stained with either PE Mouse anti-GATA3 or PE Mouse IgG1 κ Isotype control (Cat. No. 559320, not shown). When compared to the respective isotype controls, the GATA3 staining on the D10.G4.1 cell line was significantly brighter than on the 2D6 cells. Flow cytometry was performed on a BD™ FACSArray™ bioanalyzer system.
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Comparison of GATA3 expression in human T and B cell lines. Jurkat T leukemia (ATCC TIB152, shaded histogram) and Ramos Burkitt's lymphoma (ATCC CRL-1596, open histogram) were fixed with pre-warmed BD Cytofix™ buffer (Cat. No. 554655) for 10 minutes at 37ºC, permeabilized with BD™ Phosflow Perm Buffer III (Cat. No. 558050) on ice for at least 30 minutes, and then stained with either PE Mouse anti-GATA3 or PE Mouse IgG1 κ Isotype control (Cat. No. 559320, not shown). The GATA3 staining on the Jurkat cell line was significantly brighter than the isotype control on Jurkat cells, while the GATA3 staining on the Ramos cells coincided very closely to its isotype control (data not shown). Thus, GATA3 expression was detected on the T cell line but not the B cell line. Flow cytometry was performed on a BD FACSArray™ bioanalyzer system.
Comparison of GATA3 expression in mouse Th2 and Th1 cell lines. D10.G4.1 Th2 lymphoblasts (ATCC TIB-224, shaded histogram) and 2D6 Th1 clone (Ahn et al, 1998, open histogram) were fixed with pre-warmed BD Cytofix™ buffer (Cat. No. 554655) for 10 minutes at 37ºC, permeabilized with BD™ Phosflow Perm Buffer III (Cat. No. 558050) on ice for at least 30 minutes, and then stained with either PE Mouse anti-GATA3 or PE Mouse IgG1 κ Isotype control (Cat. No. 559320, not shown). When compared to the respective isotype controls, the GATA3 staining on the D10.G4.1 cell line was significantly brighter than on the 2D6 cells. Flow cytometry was performed on a BD™ FACSArray™ bioanalyzer system.
Product Details
BD Pharmingen™
Gata-3; GATA binding protein 3; GATA-binding factor 3; HDR
Human (QC Testing), Mouse (Tested in Development), Rat (Predicted)
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)
20 µl
AB_1645330
Aqueous buffered solution containing BSA and ≤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 with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.
Recommended Assay Procedures
Either BD Cytofix™ fixation buffer or BD™ Phosflow Fix Buffer I may be used for cell fixation.
Product Notices
- 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Fixation Buffer RUO
Size
100 mL
Cat No.
554655
Perm Buffer III RUO
Size
125 mL
Cat No.
558050
Fix Buffer I RUO
Size
250 mL
Cat No.
557870
PE Mouse IgG1, κ Isotype Control RUO
Size
100 Tests
Cat No.
559320
560074 Rev. 2
Antibody Details
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.
560074 Rev. 2
Format Details
PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
560074 Rev.2
Citations & References
Development References (9)
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Ahn H-J, Maruo S, Tomura M, et al. A mechanism underlying synergy between IL-12 and IFN-γ-inducing factor in enhanced production of IFN-γ. J Immunol. 1997; 159:2125-2131. (Methodology).
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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).
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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).
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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).
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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
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Usary J, Llaca V, Karaca G, et al. Mutation of GATA3 in human breast tumors. Oncogene. 2004; 23(46):7669-7678. (Biology). View Reference
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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
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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
-
van Esch H, Groenen P, Nesbit MA, et al. GATA3 haplo-insufficiency causes human HDR syndrome. Nature. 2000; 106:419-422. (Biology). View Reference
560074 Rev. 2
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
