Product Information

Contents: Alexa Fluor® 647 anti-mouse/human Gata-3 (Gata3)
Catalog Number: 51-9966
Sizes: 25 tests, 100 tests
Formulation: Phosphate buffered saline, pH 7.2, less than or equal to 0.09% NaN₃, BSA and/or gelatin for protein stabilization
Storage Conditions: Store at 4°C.
DO NOT FREEZE.
LIGHT-SENSITIVE MATERIAL.
Clone: TWAJ
Isotype: Rat IgG2b, k

Staining of CD4 (cat. 11-0041) single positive (left) or CD8 (cat. 45-0081) single positive (right) BALB/c thymocytes using Foxp3 Buffer Set (cat. 00-5523) with Alexa Fluor® 647 Rat IgG2b Isotype Control (cat. 51-4031) (blue histogram) or Alexa Fluor® 647 anti-mouse/human Gata-3 (TWAJ) (purple histogram).

Available Formats of This Product
Cat. No.	Format	Excite (nm)	Emit (nm)	Reported Applications
14-9966	Affinity Purified anti-mouse/human Gata-3 (Gata3)	N/A	N/A	WB
12-9966	Phycoerythrin (PE) anti-mouse/human Gata-3 (Gata3)	488	575	IC Flow
51-9966	Alexa Fluor® 647 anti-mouse/human Gata-3 (Gata3)	633	668	IC Flow
Flow Cytometry Product Notes: Test Sizes: To accommodate multicolor flow cytometry, eBioscience is in the process of reducing test size volumes from 20 µl to 5 µl. Please check your antibody vial for the recommended test size. Fluorochrome Replacements: eBioscience is in the process of replacing all Pacific Blue® and APC-Alexa Fluor® 750 conjugated products with eFluor™ 450 and APC-eFluor™ 780 conjugated products, respectively.

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Description

The monoclonal antibody TWAJ recognizes mouse and human Gata-3, a member of the Gata family of transcription factors. Gata-3 is a T cell specific transcription factor shown to be important for thymic development and Th2 differentiation. Expression during embryonic development is found in the central nervous system, skin, mammary glands and kidney. It’s expression is critical as Gata-3 knock-out mice survive to only day 11.5. It’s role in thymic development is essential for T cell commitment and survival. In the thymus expression is found mainly on the CD4 single positive cells. In Th2 differentiation Gata-3 binds the IL-4 promoter and the presence of Gata-3 represses T-bet expression, and hence Th1 differentiation.

Alternative splice variants have been reported especially in the MCF7 cell line. The TWAJ anti-mouse/human Gata-3 antibody will recognize both forms (50 and 45kDa) of the protein.

Staining with the TWAJ anti-mouse/human Gata-3 antibody requires the use of the Foxp3 buffers.

Applications Reported

For research use only, not for diagnostic or therapeutic use. This TWAJ antibody has been reported for use in intracellular staining followed by flow cytometric analysis.
The use of Foxp3 buffers for staining is CRITCAL.

Applications Tested

This TWAJ antibody has been pre-titrated and tested by intracellular staining usign Foxp3 buffers and flow cytometric analysis of mouse thymocytes. This can be used at 20 μl (0.125 μg) per million cells in a 100 μl total staining volume.

References

Mantel PY, Kuipers H, Boyman O, Rhyner C, Ouaked N, Rückert B, Karagiannidis C, Lambrecht BN, Hendriks RW, Crameri R, Akdis CA, Blaser K, Schmidt-Weber CB. GATA3-driven Th2 responses inhibit TGF-beta1-induced FOXP3 expression and the formation of regulatory T cells. PLoS Biol. 2007 Dec;5(12):e329.

Usary J, Llaca V, Karaca G, Presswala S, Karaca M, He X, Langerød A, Kåresen R, Oh DS, Dressler LG, Lønning PE, Strausberg RL, Chanock S, Børresen-Dale AL, Perou CM.Mutation of GATA3 in human breast tumors. Oncogene. 2004 Oct 7;23(46):7669-78.

Hernández-Hoyos G, Anderson MK, Wang C, Rothenberg EV, Alberola-Ila J. GATA-3 expression is controlled by TCR signals and regulates CD4/CD8 differentiation. Immunity. 2003 Jul;19(1):83-94.

Pai SY, Truitt ML, Ting CN, Leiden JM, Glimcher LH, Ho IC. Critical roles for transcription factor GATA-3 in thymocyte development. Immunity. 2003 Dec;19(6):863-75.

Hendriks RW, Nawijn MC, Engel JD, van Doorninck H, Grosveld F, Karis A. Expression of the transcription factor GATA-3 is required for the development of the earliest T cell progenitors and correlates with stages of cellular proliferation in the thymus. Eur J Immunol. 1999 Jun;29(6):1912-8.

Related Products

Cat. 11-0041  FITC anti-mouse CD4 (L3T4) (clone GK1.5)
Cat. 45-0081  PerCP-Cy5.5 anti-mouse CD8 (alpha subunit, CD8a, Ly-2) (clone 53-6.7)
Cat. 51-4031  Alexa Fluor® 647 Rat IgG2b Isotype Control
Cat. 00-5523  eBioscience Foxp3 Staining Buffer Set

Alexa Fluor® and Pacific Blue® are registered trademarks of and licensed under patents assigned to Molecular Probes, Inc. for research use only. This product is subject to an agreement between Molecular Probes, Inc. and eBioscience, and the manufacture, use, sale or import of this product may be subject to one or more U.S. patents, pending applications and corresponding foreign equivalents, owned by Molecular Probes, Inc. (a wholly owned subsidiary of Invitrogen Corp). The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product for life science research or as an ASR. The buyer cannot use this product for manufacturing or for any other screening (specifically including use in combination with microarrays or High Content Screening) or testing purpose, other than as an ASR. For information on purchasing a license to this product for purposes other than life science research or use as an ASR, contact Molecular Probes, Inc.