Flow cytometric analysis of GATA3 expression in Human 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™ RB705 Mouse Anti-GATA3 antibody (Cat. No. 570618/570706). The fluorescence histograms showing GATA3 expression were derived from gated events with the forward and side light-scatter characteristics of intact lymphoid cells. Flow cytometry and data analysis were performed using a LSRFortessa™ X-20 Flow Cytometer System and FlowJo™ Software.
Flow cytometric analysis of GATA3 expression in Mouse D10.G4.1 cells. Cells from the Mouse 2D6 (Ahn et al, 1998; dashed line histogram) and D10.G4.1 (T Helper ATCC® TIB-224™; 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™ RB705 Mouse Anti-GATA3 antibody (Cat. No. 570618/570706). The fluorescence histograms showing GATA3 expression were derived from gated events with the forward and side light-scatter characteristics of intact lymphoid cells. Flow cytometry and data analysis were performed using a LSRFortessa™ X-20 Flow Cytometer System and FlowJo™ Software.
Flow cytometric analysis of GATA3 expression in Human 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™ RB705 Mouse Anti-GATA3 antibody (Cat. No. 570618/570706). The fluorescence histograms showing GATA3 expression were derived from gated events with the forward and side light-scatter characteristics of intact lymphoid cells. Flow cytometry and data analysis were performed using a LSRFortessa™ X-20 Flow Cytometer System and FlowJo™ Software.
Flow cytometric analysis of GATA3 expression in Mouse D10.G4.1 cells. Cells from the Mouse 2D6 (Ahn et al, 1998; dashed line histogram) and D10.G4.1 (T Helper ATCC® TIB-224™; 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™ RB705 Mouse Anti-GATA3 antibody (Cat. No. 570618/570706). The fluorescence histograms showing GATA3 expression were derived from gated events with the forward and side light-scatter characteristics of intact lymphoid cells. Flow cytometry and data analysis were performed using a LSRFortessa™ X-20 Flow Cytometer System and FlowJo™ Software.
Product Details
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/test
2625, 14462
Aqueous buffered solution containing ≤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 to the dye under optimum conditions and unreacted dye was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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
- When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
- Tandem fluorochromes contain both an energy donor and an energy acceptor. 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 the donor may be observed. Additionally, multi-laser cytometers may directly excite both the donor and acceptor fluorochromes. Therefore, we recommend for every tandem conjugate, a matched individual single-stain control be acquired for generating a compensation or spectral unmixing matrix.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
- For U.S. patents that may apply, see bd.com/patents.
570618 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.
570618 Rev. 1
Format Details
RB705
The BD Horizon RealBlue™ 705 (RB705) Dye is part of the BD® family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 707-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RB705 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), RB705 can be used as an alternative to PerCP-Cy5.5 or BB700 and we recommend using an optical filter centered near 710-nm (e.g., a 695/40 or 710/50-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PerCP-Cy5.5. RB705 is on average brighter than PerCP-Cy5.5 and BB700, and has minimal spillover into Yellow-Green detectors.
RB705
Blue 488 nm
498 nm
707 nm
570618 Rev.1
Citations & References
View product citations for antibody "570618" on CiteAb
Development References (8)
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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
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Hayatsu N, Miyao T, Tachibana M, et al. Analyses of a Mutant Foxp3 Allele Reveal BATF as a Critical Transcription Factor in the Differentiation and Accumulation of Tissue Regulatory T Cells.. Immunity. 2017; 47(2):268-283.e9. (Clone-specific: Intracellular Staining/Flow Cytometry). View Reference
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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
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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
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Rudra D, deRoos P, Chaudhry A, et al. Transcription factor Foxp3 and its protein partners form a complex regulatory network.. Nat Immunol. 2012; 13(10):1010-9. (Clone-specific: Immunoprecipitation, Intracellular Staining/Flow Cytometry, Western blot). 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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Yu JC, Lin G, Field JJ, Linden J. Induction of antiinflammatory purinergic signaling in activated human iNKT cells.. JCI Insight. 2018; 3(17):e9195491954. (Clone-specific: Intracellular Staining/Flow Cytometry). 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
570618 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.
Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.