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-Cy™7 Mouse anti-GATA3 or PE-Cy™7 Mouse IgG1 κ Isotype Control (clone MOPC-21, Cat. No. 557646, 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 FACSCalibur™ flow cytometry 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-Cy™7 Mouse anti-GATA3 or PE-Cy™7 Mouse IgG1 κ Isotype Control (clone MOPC-21, Cat. No. 557646, 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 FACSCalibur™ flow cytometry 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-Cy™7 Mouse anti-GATA3 or PE-Cy™7 Mouse IgG1 κ Isotype Control (clone MOPC-21, Cat. No. 557646, 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 FACSCalibur™ flow cytometry 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-Cy™7 Mouse anti-GATA3 or PE-Cy™7 Mouse IgG1 κ Isotype Control (clone MOPC-21, Cat. No. 557646, 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 FACSCalibur™ flow cytometry system.
Product Details
BD Pharmingen™
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)
20 µl
AB_1645544
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO
Preparation And Storage
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with PE-Cy7 under optimum conditions, and unconjugated antibody and free PE-Cy7 were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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).
- PE-Cy7 is a tandem fluorochrome composed of R-phycoerythrin (PE), which is excited by 488-nm light and serves as an energy donor, coupled to the cyanine dye Cy7, which acts as an energy acceptor and fluoresces maximally at 780 nm. PE-Cy7 tandem fluorochrome emission is collected in a detector for fluorescence wavelengths of 750 nm and higher. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from PE may be observed. Therefore, we recommend that individual compensation controls be performed for every PE-Cy7 conjugate. PE-Cy7 is optimized for use with a single argon ion laser emitting 488-nm light, and there is no significant overlap between PE-Cy7 and FITC emission spectra. When using dual-laser cytometers, which may directly excite both PE and Cy7, we recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
- Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
- Warning: Some APC-Cy7 and PE-Cy7 conjugates show changes in their emission spectrum with prolonged exposure to formaldehyde. If you are unable to analyze fixed samples within four hours, we recommend that you use BD™ Stabilizing Fixative (Cat. No. 338036).
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Cy is a trademark of Amersham Biosciences Limited. This conjugated product is sold under license to the following patents: US Patent Nos. 5,486,616; 5,569,587; 5,569,766; 5,627,027.
- This product is subject to proprietary rights of Amersham Biosciences Corp. and Carnegie Mellon University and made and sold under license from Amersham Biosciences Corp. This product is licensed for sale only for research. It is not licensed for any other use. If you require a commercial license to use this product and do not have one return this material, unopened to BD Biosciences, 10975 Torreyana Rd, San Diego, CA 92121 and any money paid for the material will be refunded.
- 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.
Companion Products
Fixation Buffer RUO
Size
100 mL
Cat No.
554655
Perm Buffer III RUO
Size
125 mL
Cat No.
558050
PE-Cy™7 Mouse IgG1 κ Isotype Control RUO
Size
100 Tests
Cat No.
557646
560405 Rev. 1
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.
560405 Rev. 1
Format Details
PE-Cy7
PE-Cy7 dye is a part of the BD PE family of dyes. This tandem fluorochrome is comprised of a R-Phycoerythrin (PE) donor that has excitation maxima (Ex Max) of 496-nm and 566-nm and an acceptor dye, Cy™7, with an emission maximum (Em Max) at 781-nm. PE can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and detected using an optical filter centered near 781 nm (e.g., a 760/60-nm bandpass filter). The donor dye can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and the acceptor dye can be excited by the Red (627–640-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
PE-Cy7
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
781 nm
560405 Rev.1
Citations & References
Development References (9)
-
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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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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Tong Q, Hotamisligil GS. Cell fate in the mammary gland. Nature. 2008; 556:724-726. (Biology).
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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
560405 Rev. 1
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.
