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 Alexa Fluor® 488 Mouse anti-GATA3 (Cat. No. 560163/560077) or Alexa Fluor® 488 Mouse IgG1 κ Isotype control (Cat. No. 557782, 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™ LSR II 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 Alexa Fluor® 488 Mouse anti-GATA3 or Alexa Fluor® 488 Mouse IgG1 κ Isotype control (Cat. No. 557782, 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™ LSR II 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 Alexa Fluor® 488 Mouse anti-GATA3 (Cat. No. 560163/560077) or Alexa Fluor® 488 Mouse IgG1 κ Isotype control (Cat. No. 557782, 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™ LSR II 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 Alexa Fluor® 488 Mouse anti-GATA3 or Alexa Fluor® 488 Mouse IgG1 κ Isotype control (Cat. No. 557782, 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™ LSR II flow cytometry 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_1645302
Aqueous buffered solution containing BSA, protein stabilizer, 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 to Alexa Fluor® 488 under optimum conditions, and unreacted Alexa Fluor® 488 was 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- 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
Fix Buffer I RUO
Size
250 mL
Cat No.
557870
Perm Buffer III RUO
Size
125 mL
Cat No.
558050
Alexa Fluor® 488 Mouse IgG1 κ Isotype Control RUO
Size
50 Tests
Cat No.
557782
560163 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.
560163 Rev. 2
Format Details
Alexa Fluor™ 488
Alexa Fluor™ 488 Dye is part of the BD blue family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 494-nm and an emission maximum (Em Max) at 517-nm. Alexa Fluor™ 488 is designed to be excited by the Blue laser (488 nm) and detected using an optical filter centered near 520-nm (e.g., a 530/30-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 488
Blue 488 nm
494 nm
517 nm
560163 Rev.2
Citations & References
View product citations for antibody "560163" on CiteAb
Development References (10)
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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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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
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van Esch H, Groenen P, Nesbit MA, et al. GATA3 haplo-insufficiency causes human HDR syndrome. Nature. 2000; 106:419-422. (Biology). View Reference
560163 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.