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Preparation And Storage
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
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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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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.