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BD Horizon™ RB780 Rat Anti-Mouse Zbtb7b (ThPok)
Clone T43-94 (RUO)
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.
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.
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.
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.
Product Details
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
AB_3684475
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
- 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
- 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.
- 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
Companion Products
Stain Buffer (FBS) RUO
Size
500 ML
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 ML
Cat No.
554657
Transcription Factor Buffer Set RUO
Size
100 Tests
Cat No.
562574
RB780 Rat IgG2b, κ Isotype Control RUO
Size
50 µg
Cat No.
568699
BV480 Rat Anti-Mouse CD4 RUO
Size
50 µg
Cat No.
565634
568697 Rev. 2
Antibody Details
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.
568697 Rev. 2
Format Details
RB780
The BD Horizon RealBlue™ 780 (RB780) Dye is part of the BD family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 781-nm. Driven by BD innovation, RB780 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), RB780 can be used as an alternative to PE-Cy7 and we recommend using an optical filter centered near 780-nm (eg, a 780/60-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PE-Cy7. RB780 is on average brighter than PE-Cy7 and has minimal spillover into Yellow-Green detectors.
RB780
498 nm
781 nm
568697 Rev.2
Citations & References
View product citations for antibody "568697" on CiteAb
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
568697 Rev. 2
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.