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 important for thymic development and Th2 differentiation. Expression during embryonic development is found in the central nervous system, skin, mammary glands and kidney. During development, the expression of Gata-3 is essential as homozygous knock-out of Gata-3 is embryonic lethal. The Gata-3 is also essential for T cell commitment and survival. In the thymus, expression is found mainly on the CD4 single positive cells. During Th2 differentiation, Gata-3 binds to the IL-4 promoter as well as represses the expression of T-bet, thus inhibiting Th1 differentiation.
Alternative splice variants have been reported especially in the MCF7 cell line. The TWAJ Human/Mouse Gata-3 antibody will recognize both forms (50 and 45 kDa) of the protein.
Staining with the TWAJ Human/Mouse Gata-3 antibody requires the use of the Foxp3/Transcription Factor Staining Buffer Set.(cat 00-5523)
Crossreactivity in rhesus monkeys has been published.
Applications Reported: This TWAJ antibody has been reported for use in intracellular staining followed by flow cytometric analysis.
Applications Tested: This TWAJ antibody has been pre-titrated and tested by intracellular staining and flow cytometric analysis of mouse thymocytes using the Foxp3/Transcription Factor Staining Buffer Set (cat. 00-5523) and protocol. Please refer to Best Protocols: Protocol B: One step protocol for (nuclear) intracellular proteins located under the Resources Tab online. This can be used at 5 µL (0.06 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.
PerCP-eFluor® 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5. We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative. Please make sure that your instrument is capable of detecting this fluorochrome.
Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.
Fixation: Samples can be stored in IC Fixation Buffer (cat. 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.
Excitation: 488 nm; Emission: 710 nm; Laser: Blue Laser.
Filtration: 0.2 µm post-manufacturing filtered.
The genes for all 4 subunits of the T-cell antigen receptor (alpha, beta, gamma and delta) are controlled by distinct enhancers and their enhancer-binding proteins. Marine and Winoto (1991) identified a common TCR regulatory element by demonstrating binding of the enhancer-binding protein GATA3 to the enhancer elements of all 4 TCR genes. GATA3 had been shown in the chicken to be an enhancer-binding protein containing a zinc finger domain. GATA3 mRNA was demonstrated by Northern blot analysis in T cells but not in B cells or macrophages. GATA3 is abundantly expressed in the T-lymphocyte lineage and is thought to participate in T-cell receptor gene activation through binding to enhancers. Labastie et al. (1994) cloned the human gene and the 5-prime end of the mouse gene. The human gene comprises 6 exons distributed over 17 kb of DNA. Its 2 zinc fingers are encoded by 2 separate exons highly conserved with those of GATA1, but no other structural homologies between the 2 genes could be found.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Protein Aliases: GATA binding factor-3; GATA binding protein 3; GATA-binding factor 3; GATA-binding protein 3; HDR; MGC2346; MGC5199; MGC5445; Trans-acting T-cell-specific transcription factor GATA-3
Gene Aliases: EGK_19445; Gata-3; GATA3; HDR; HDRS; jal