The antibody was purified by affinity chromatography.
Storage & Handling:
The antibody solution should be stored undiluted between 2°C and 8°C.
WB, ICFC - Quality tested IF, IP - Reported in the literature
Additional reported applications (for the relevant formats) include: immunoprecipitation2 and immunofluorescence microscopy3.
NOTE: For flow cytometric staining with this clone, True-Nuclear™ Transcription Factor Buffer Set offers improved staining and is highly recommended over the Foxp3 Fix/Perm Buffer Set and the Nuclear Factor Fixation and Permeabilization Buffer Set .
Each lot of this antibody is quality control tested by True-Nuclear™ Transcription Factor Staining Protocol. For Western blotting, the suggested use is 1 to 2 ug per ml. For flow cytometric staining, the suggested use of this reagent is 1.0 µg per million cells in a staining volume of 100 µl. It is recommended that the reagent be titrated for optimal performance for each application.
1. Szabo SJ, et al. 2000. Cell 100:655. (ICFC, WB) 2. Hwang ES, et al. 2005. J. Exp. Med. 202:1289. (ICFC, WB, IP) 3. Neurath MF, et al. 2002. J. Exp. Med. 195:1129. (IF) 4. Hsieh CY, et al. 2012. J Pharmacol Exp. 343:125. PubMed.
Total cell lysate from PBMC (lane 1, 15 µg) and PBMC treated with 5 µg/mL CD3 and 2 µg/mL CD28 (lane 2, 15 µg) were resolved by electrophoresis (4-12% Bis-Tris), transferred to nitrocellulose, and probed with purified anti-T-bet antibody (clone 4B10). Proteins were visualized using an HRP Goat anti-mouse IgG Antibody and chemiluminescence detection. Direct-Blot™ HRP anti-β-actin antibody (clone 2F1-1) was used as a loading control.
Human peripheral blood lymphocytes were surface stained with CD3 (clone OKT3) APC and then treated with True-Nuclear™ Transcription Factor Buffer Set. The cells were then stained with purified T-bet (clone 4B10) followed by anti-mouse IgG1 PE.
T-bet, also known as T-box transcription factor T-bet, is considered to be a "master regulator" of Th1 lymphoid development controlling the production of the cytokine IFN-γ. T-bet is widely expressed in hematopoietic cells including stem cells, NK cells, B cells, and T cells. T-bet is critical for the control of microbial pathogens, and knockout animals show multiple physiologic and inflammatory features characteristic of asthma. T-bet expression is optimally observed after IL-12 stimulation and can be suppressed by addition of the Th2 cytokine IL-4 or neutralization of IL-12.
T-box expressed in T cells, T box 21, TBLYM
T-box transcription factor, approximately 58 kD.
Nuclear; expressed in T cells, hematopoietic stem cells, NK cells, B cells, lung, spleen.
Th1-specific T-box transcription factor controlling expression of the hallmark Th1 cytokine, interferon gamma (IFN-γ). T-bet expression is critical for the control of microbial pathogens.
1. Szabo SJ, et al. 2000. Cell 100:655. 2. Szabo SJ, et al. 2002. Science 295:338. 3. Finotto S, et al. 2002. Science 295:336. 4. Mullen AC, et al. 2001. Science 292:1907.
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