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RY586 Rat Anti-Mouse Zbtb7b
RY586 Rat Anti-Mouse Zbtb7b
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™ Rat Anti-Mouse CD4 antibody (Cat. No. 562891) and with either BD Horizon™ RY586 Rat IgG2b, κ Isotype Control (Cat. No. 568160; Left Plot) or BD Horizon™ RY586 Rat Anti-Mouse Zbtb7b (ThPok) antibody (Cat. No. 568128/568129; Right Plot) at 0.03 µg/test. The bivariate 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 LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
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™ Rat Anti-Mouse CD4 antibody (Cat. No. 562891) and with either BD Horizon™ RY586 Rat IgG2b, κ Isotype Control (Cat. No. 568160; Left Plot) or BD Horizon™ RY586 Rat Anti-Mouse Zbtb7b (ThPok) antibody (Cat. No. 568128/568129; Right Plot) at 0.03 µg/test. The bivariate 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 LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
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BD Horizon™
Zbtb7b; ZBT7B; Th-POK; Thpok; c-Krox; Zfp-67; Zfp67
Mouse (QC Testing)
Rat WI, also known as Wistar (outbred) IgG2b, κ
Mouse Zbtb7b
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unreacted dye was removed.

Recommended Assay Procedures

BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. 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.
  3. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  4. An isotype control should be used at the same concentration as the antibody of interest.
  5. CF™ is a trademark of Biotium, Inc.
  6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  7. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
568129 Rev. 2
Antibody Details
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T43-94

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.

The BD Horizon RealYellow™ 586 (RY586) Dye is part of the BD family of yellow-green dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 565-nm and an emission maximum (Em Max) at 586-nm. Driven by BD innovation, RY586 can be used on both spectral and conventional cytometers and is designed to be excited by the Yellow-Green laser (561-nm) with minimal excitation by the 488-nm Blue laser. For conventional instruments equipped with a Yellow-Green laser (561-nm), RY586 can be used as an alternative to PE and we recommend using an optical filter centered near 586-nm (eg, a 586/15-nm bandpass filter). For spectral instruments equipped with a Yellow-Green laser (561-nm), it can be used in conjunction with PE. Compared to PE, RY586 is similar in brightness, minimal spillover into Blue detectors, and increased spillover into the 610/20-nm (PE-CF594) detector. Please ensure that your instrument configuration (lasers and optical filters) is appropriate for this dye.

568129 Rev. 2
Format Details
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RY586
The BD Horizon RealYellow™ 586 (RY586) Dye is part of the BD family of yellow-green dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 565-nm and an emission maximum (Em Max) at 586-nm. Driven by BD innovation, RY586 can be used on both spectral and conventional cytometers and is designed to be excited by the Yellow-Green laser (561-nm) with minimal excitation by the 488-nm Blue laser. For conventional instruments equipped with a Yellow-Green laser (561-nm), RY586 can be used as an alternative to PE and we recommend using an optical filter centered near 586-nm (eg, a 586/15-nm bandpass filter). For spectral instruments equipped with a Yellow-Green laser (561-nm), it can be used in conjunction with PE. Compared to PE, RY586 is similar in brightness, minimal spillover into Blue detectors, and increased spillover into the 610/20-nm (PE-CF594) detector. Please ensure that your instrument configuration (lasers and optical filters) is appropriate for this dye.
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RY586
Yellow-Green 561 nm
564 nm
586 nm
568129 Rev.2
Citations & References
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View product citations for antibody "568129" on CiteAb

Development References (7)

  1. 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. (Clone-specific: Flow cytometry). View Reference
  2. Kappes DJ. Expanding roles for ThPOK in thymic development. Immunol Rev. 2010; 238(1):182-194. (Biology). View Reference
  3. 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
  4. 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
  5. 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
  6. Taniuchi I, Ellmeier W. Transcriptional and epigenetic regulation of CD4/CD8 lineage choice. Adv Immunol. 2011; 110:71-110. (Biology). View Reference
  7. 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. (Clone-specific: Flow cytometry). View Reference
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568129 Rev. 2

 

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