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Two-color flow cytometric analysis of Zbtb7b (ThPok) expression in Mouse thymocytes. Mouse thymocytes were fixed and permeabilized with BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725) and stained with BD Horizon™ BV480 Rat Anti-Mouse CD4 antibody (Cat. No. 565634) and with either BD Horizon™ RB780 Rat IgG2b, κ Isotype Control (Cat. No. 568699; Left Plot) or BD Horizon™ RB780 Rat Anti-Mouse Zbtb7b (ThPok) antibody (Cat. No. 568696/568697; Right Plot). The two-parameter pseudocolor density plot showing the correlated expression of Zbtb7b (ThPok) [or Ig Isotype control staining] versus CD4 was derived from gated events with the forward and side light-scatter characteristics of intact thymocytes. Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 Cell Analyzer System and FlowJo™ Software.
BD Horizon™ RB780 Rat Anti-Mouse Zbtb7b (ThPok)
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Preparation And Storage
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- When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
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The T43-94 monoclonal antibody specifically recognizes mouse Zinc finger and BTB domain-containing protein 7B (Zbtb7b) which is also known as, T-helper-inducing POZ/Krueppel-like factor (Th-POK), Krueppel-related zinc finger protein cKrox (cKrox), or Zinc finger protein 67 (Zfp67). Zbtb7b is a Zn2+ finger-containing transcription factor that acts as a transcriptional repressor or activator in a promoter-dependent manner. Zbtb7b plays a role in thymic selection of T cells and is expressed in CD4+CD8+ thymocytes and mature CD4+CD8- thymocytes and T cells. It is not expressed by mature CD4-CD8+ thymocytes and T cells. It indirectly increases the expression of CD4 in developing T cells by antagonizing Runx-3-mediated CD4 repression. Zbtb7b expression has also been found in NKT and gamma/delta T cells. Zbtb7b plays a role in transcriptional repression of collagen gene expression as well.
Development References (8)
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Chopp LB, Gopalan V, Ciucci T, et al. An Integrated Epigenomic and Transcriptomic Map of Mouse and Human αβ T Cell Development.. Immunity. 2020; 53(6):1182-1201.e8. (Clone-specific: Flow cytometry). View Reference
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Ciucci T, Vacchio MS, Gao Y, et al. The Emergence and Functional Fitness of Memory CD4+ T Cells Require the Transcription Factor Thpok.. Immunity. 2019; 50(1):91-105.e4. (Clone-specific: Flow cytometry). View Reference
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Kappes DJ. Expanding roles for ThPOK in thymic development. Immunol Rev. 2010; 238(1):182-194. (Biology). View Reference
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Luckey MA, Kimura MY, Waickman AT, Feigenbaum L, Singer A, Park JH. The transcription factor ThPOK suppresses Runx3 and imposes CD4(+) lineage fate by inducing the SOCS suppressors of cytokine signaling. Nat Immunol. Nat Immunol. 2014; 15(7):638-645. (Biology). View Reference
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Mittal P, Abblett R, Ryan JM, et al. An Immunotherapeutic CD137 Agonist Releases Eomesodermin from ThPOK Repression in CD4 T Cells.. J Immunol. 2018; 200(4):1513-1526. (Clone-specific: Flow cytometry). View Reference
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Reis BS, Rogoz A, Costa-Pinto FA, Taniuchi I, Mucida D. Mutual expression of the transcription factors Runx3 and ThPOK regulates intestinal CD4(+) T cell immunity. Nat Immunol. 2013; 14(3):271-280. (Biology). View Reference
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Taniuchi I, Ellmeier W. Transcriptional and epigenetic regulation of CD4/CD8 lineage choice. Adv Immunol. 2011; 110:71-110. (Biology). View Reference
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Vacchio MS, Ciucci T, Gao Y, et al. A Thpok-Directed Transcriptional Circuitry Promotes Bcl6 and Maf Expression to Orchestrate T Follicular Helper Differentiation.. Immunity. 2019; 51(3):465-478.e6. (Clone-specific: Flow cytometry). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.