Intracellular Staining by Flow Cytometry
|Detection of T‑bet/TBX21 in Jurkat Human Cell Line by Flow Cytometry. Jurkat human acute T cell leukemia cell line was stained with Human T‑bet/TBX21 Monoclonal Antibody (Catalog # MAB5385, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG F(ab')2 Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with ice-cold methanol.|
|Detection of Human T-bet/TBX21 by Western Blot. Western blot shows lysates of Raji human Burkitt's lymphoma cell line and Daudi human Burkitt's lymphoma cell line. PVDF membrane was probed with 2 µg/mL of Human T-bet/TBX21 Monoclonal Antibody (Catalog # MAB5385) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for T-bet/TBX21 at approximately 55 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.|
T-box expressed in T cells (T-bet), also known as T-box transcription factor TBX21, is a 62 kDa member of the T-box family of transcription factors and the Tbr1 subfamily. Human T-bet is 535 amino acids in length and contains a T-box DNA binding domain (aa 136-327). Human T-bet shares 88% aa sequence identity with mouse T-bet. T-bet is a nuclear protein highly apparent in Th1-cells. Northern blot analysis revealed that it is also expressed in lung, thymus and spleen. Functionally, T-bet controls the expression of the Th1 cytokine, IFN gamma, and initiates Th1 lineage development from naïve Th precursor cells by both activating Th1 genetic programs and by repressing the opposing Th2 programs.
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