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566942_2.png

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 FITC Rat Anti-Mouse CD4 antibody (Cat. No. 553046/553047/561835) and either BD Horizon™ R718 Rat IgG2b, κ Isotype Control (Cat. No. 566943; Left Plot) or BD Horizon™ R718 Rat Anti-Mouse Zbtb7b (ThPok) antibody (Cat. No. 566942; Right Plot). A 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 LSRFortessa™ Flow Cytometer System and FlowJo™ software.

566942Image1.png

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 FITC Rat Anti-Mouse CD4 antibody (Cat. No. 553046/553047/561835) and either BD Horizon™ R718 Rat IgG2b, κ Isotype Control (Cat. No. 566943; Left Plot) or BD Horizon™ R718 Rat Anti-Mouse Zbtb7b (ThPok) antibody (Cat. No. 566942; Right Plot). A 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 LSRFortessa™ Flow Cytometer System and FlowJo™ software.

566942_2.png 566942Image1.png Blue-Cap-Red-Label.jpg

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 FITC Rat Anti-Mouse CD4 antibody (Cat. No. 553046/553047/561835) and either BD Horizon™ R718 Rat IgG2b, κ Isotype Control (Cat. No. 566943; Left Plot) or BD Horizon™ R718 Rat Anti-Mouse Zbtb7b (ThPok) antibody (Cat. No. 566942; Right Plot). A 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 LSRFortessa™ Flow Cytometer 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 FITC Rat Anti-Mouse CD4 antibody (Cat. No. 553046/553047/561835) and either BD Horizon™ R718 Rat IgG2b, κ Isotype Control (Cat. No. 566943; Left Plot) or BD Horizon™ R718 Rat Anti-Mouse Zbtb7b (ThPok) antibody (Cat. No. 566942; Right Plot). A 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 LSRFortessa™ Flow Cytometer System and FlowJo™ software.

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
AB_2869959
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.
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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 CompBead to ensure that BD CompBeads are appropriate for your specific cellular application.

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Product Notices

  1. 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).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. 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.
  4. Alexa Fluor® is a registered trademark of Life Technologies Corporation.
  5. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  7. This product is provided under an Agreement between BIOTIUM and BD Biosciences. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
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566942 Rev. 3
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 antibody was conjugated to BD Horizon Red 718, which has been developed exclusively for BD Biosciences as a better alternative to Alexa Fluor® 700. BD Horizon Red 718 can be excited by the red laser (628 – 640 nm) and, with an Em Max around 718 nm, it can be detected using a 730/45 nm filter. Due to similar excitation and emission properties, we do not recommend using R718 in combination with APC-R700 or Alexa Fluor® 700.

566942 Rev. 3
Format Details
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R718
The BD Horizon™ Red 718 (R718) Dye is part of the BD red family of dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 695-nm and an emission maximum (Em Max) at 718-nm. Driven by BD innovation, R718 is designed to be excited by the red laser (627–640-nm) and detected using an optical filter centered near 720-nm (e.g., a 720/40-nm bandpass filter). R718 is a brighter alternative to Alexa Fluor™ 700. R718 is also a bright small molecule alternative to APC-R700 with lower spread into the APC detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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R718
695 nm
718 nm
566942 Rev.3
Citations & References
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View product citations for antibody "566942" 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.e4. (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.e6. (Clone-specific: Flow cytometry). View Reference
View All (7) View Less
566942 Rev. 3

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.