BD Pharmingen™ Purified Mouse Anti-Mouse Pre-T Cell Receptor α Chain
克隆 2F5 (RUO)
Multicolor analysis of the expression of pTα on thymocyte subsets. C57BL/6 thymocytes were stained with either purified mAb 2F5 (top right panel, bottom panels, (C-F) open histograms) or purified mouse IgG1, κ isotype control MOPC-31C (Cat. No. 557273, filled histograms) in the presence of Mouse Fc Block™ (purified anti-mouse CD16/CD32 mAb, Cat. No. 553141/533142), followed by biotinylated anti-mouse IgG1 mAb A85-1 (Cat. No. 553441), then Streptavidin-PE (Cat. No. 554061). FITC-conjugated anti-mouse CD44 mAb IM7 (Cat. No. 553133) and APC-conjugated anti-mouse CD25 mAb PC61 (Cat. No. 557192) were added in a fourth step. PerCP-conjugated anti-mouse CD4 mAb RM4-5 (Cat. No. 553052) and anti-mouse CD8a mAb 53-6.7 (Cat. No. 553036) were included in the fifth staining step. For data analysis, CD4- CD8a- (DN) thymocytes were gated as indicated in the top left panel. The expression pattern of CD44 and CD25 in that DN population is displayed in the top middle panel. The remaining panels display histograms of pTα expression on the gated subpopulations of DN thymocytes: Panel C, CD44-/lo CD25hi; Panel D, CD44- CD25lo; Panel E, CD44- CD25-; and Panel F, CD44+ CD25-. Flow cytometry was performed on a BD FACSCalibur™ System (BD Biosciences, San Jose, CA).
Multicolor analysis of the expression of pTα on thymocyte subsets. C57BL/6 thymocytes were stained with either purified mAb 2F5 (top right panel, bottom panels, (C-F) open histograms) or purified mouse IgG1, κ isotype control MOPC-31C (Cat. No. 557273, filled histograms) in the presence of Mouse Fc Block™ (purified anti-mouse CD16/CD32 mAb, Cat. No. 553141/533142), followed by biotinylated anti-mouse IgG1 mAb A85-1 (Cat. No. 553441), then Streptavidin-PE (Cat. No. 554061). FITC-conjugated anti-mouse CD44 mAb IM7 (Cat. No. 553133) and APC-conjugated anti-mouse CD25 mAb PC61 (Cat. No. 557192) were added in a fourth step. PerCP-conjugated anti-mouse CD4 mAb RM4-5 (Cat. No. 553052) and anti-mouse CD8a mAb 53-6.7 (Cat. No. 553036) were included in the fifth staining step. For data analysis, CD4- CD8a- (DN) thymocytes were gated as indicated in the top left panel. The expression pattern of CD44 and CD25 in that DN population is displayed in the top middle panel. The remaining panels display histograms of pTα expression on the gated subpopulations of DN thymocytes: Panel C, CD44-/lo CD25hi; Panel D, CD44- CD25lo; Panel E, CD44- CD25-; and Panel F, CD44+ CD25-. Flow cytometry was performed on a BD FACSCalibur™ System (BD Biosciences, San Jose, CA).
准备和存储
推荐的实验流程
This antibody has been tested by cell-surface immunofluorescent staining ( ≤8µg/million cells) with flow cytometric analysis to assure specificity and reactivity. Since the pTα is expressed at low density on the surface of immature thymocytes, we recommend the use of Mouse Fc Block™ (anti-mouse CD16/CD32 mAb 2.4G2, Cat. No. 553141/553142) and amplification of the staining intensity through the use of a biotinylated second-step antibody, such as anti-mouse IgG1 mAb A85-1 (Cat. No. 553441), followed by a "bright" third-step reagent, such as Streptavidin-PE (Cat. No. 554061).
商品通知
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
配套商品
At very early stages in the intrathymic differentiation of T lymphocytes which will bear αβ T Cell Receptors (TCR), the TCR β chain forms a heterodimer with the pre-TCR α chain (pTα) to form the pre-TCR. This pre-TCR associates with the signal-transducing CD3 complex and controls the survival and proliferation of CD4- CD8- (DN) thymocytes, plays a role in allelic exclusion of the TCR β chain, directly or indirectly regulates TCR δ chain expression, and is eventually replaced by the TCR α chain as thymocytes mature. The 2F5 monoclonal antibody specifically recognizes the PTα on the surface of CD44- CD25-/lo DN thymocytes.
研发参考 (7)
-
Aifantis I, Azogui O, Feinberg J, Saint-Ruf C, Buer J, von Boehmer H. On the role of the pre-T cell receptor in alphabeta versus gammadelta T lineage commitment. Immunity. 1998; 9(5):649-688. (Immunogen: Immunoprecipitation). 查看参考
-
Aifantis I, Feinberg J, Fehling HJ, Di Santo JP, von Boehmer H. Early T cell receptor beta gene expression is regulated by the pre-T cell receptor-CD3 complex. J Exp Med. 1999; 190(1):141-144. (Biology). 查看参考
-
Aifantis I, Pivniouk VI, Gartner F, et al. Allelic exclusion of the T cell receptor beta locus requires the SH2 domain-containing leukocyte protein (SLP)-76 adaptor protein. J Exp Med. 1999; 190(8):1093-1102. (Biology). 查看参考
-
Mancini S, Candeias SM, Fehling HJ, von Boehmer H, Jouvin-Marche E, Marche PN. TCR alpha-chain repertoire in pTalpha-deficient mice is diverse and developmentally regulated: implications for pre-TCR functions and TCRA gene rearrangement. J Immunol. 1999; 163(11):6053-6059. (Biology). 查看参考
-
Wiest DL, Berger MA, Carleton M. Control of early thymocyte development by the pre-T cell receptor complex: A receptor without a ligand. Semin Immunol. 1999; 11(4):251-262. (Biology). 查看参考
-
von Boehmer H, Aifantis I, Feinberg J, et al. Pleiotropic changes controlled by the pre-T-cell receptor. Curr Opin Immunol. 1999; 11(2):135-142. (Biology). 查看参考
-
von Boehmer H, Fehling HJ. Structure and function of the pre-T cell receptor. Annu Rev Immunol. 1997; 15:433-452. (Biology). 查看参考
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.