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Two-color flow cytometric analysis of TCF-7/TCF-1 expression by human lymphocytes or mouse splenocytes. Upper Plots - Human peripheral blood mononuclear cells (PBMC) were fixed and permeabilized with the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725). The cells were then stained with APC Mouse Anti-Human CD4 antibody (Cat. No. 555349/561840/561841) and either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; Left Plot) or BD Horizon BV421 Mouse Anti-TCF-7/TCF-1 antibody (Cat. No. 566692; Right Plot) at 0.125 μg/test. Lower Plots - Mouse splenic leucocytes were similarly fixed and permeabilized. The cells were then stained with APC Rat Anti-Mouse CD4 antibody (Cat. No. 553051/561091) and either BD Horizon BV421 Mouse IgG1, κ Isotype Control (Left Plot) or BD Horizon BV421 Mouse Anti-TCF-7/TCF-1 antibody (Right Plot) at 0.125 μg/test. Bivariate pseudocolor density plots showing the correlated expression of CD4 versus TCF-7/TCF-1 (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. 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.
BD Horizon™ BV421 Mouse Anti-TCF-7/TCF-1
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
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.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
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Companion Products
The S33-966 monoclonal antibody specifically binds to TCF-7 (Transcription factor 7) which is also known as TCF-1 (T-cell factor 1). TCF-7 is a member of the High Mobility Group (HMG) DNA binding protein family of transcription factors. TCF-7 is expressed in thymocytes, T lymphocytes and proliferating epithelial cells. TCF-7 is expressed in the earliest T cell precursors and serves as a critical transcriptional regulator that is involved in the differentiation of thymocytes to become mature T lymphocytes. TCF-7 is necessary for the survival of immature CD4+CD8+ thymocytes. TCF-7 can also promote the differentiation of mature peripheral T cells to become Th2-like cells. TCF-7 may also function as a transcriptional repressor of certain target genes including CTNNB1 and TCF7L2. The S33-966 antibody crossreacts with human TCF-7.
Development References (5)
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Chen Z, Ji Z, Ngiow SF, et al. TCF-1-Centered Transcriptional Network Drives an Effector versus Exhausted CD8 T Cell-Fate Decision.. Immunity. 2019; 51(5):840-855.e5. (Clone-specific: Flow cytometry). View Reference
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Germar K, Dose M, Konstantinou T, et al. T-cell factor 1 is a gatekeeper for T-cell specification in response to Notch signaling. Proc Natl Acad Sci U S A. 2011; 108(50):20060-20065. (Biology). View Reference
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Johnson JL, Georgakilas G, Petrovic J, et al. Lineage-Determining Transcription Factor TCF-1 Initiates the Epigenetic Identity of T Cells.. Immunity. 2018; 48(2):243-257.e10. (Clone-specific: Flow cytometry). View Reference
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Weber BN, Chi AW, Chavez A, et al. A critical role for TCF-1 in T-lineage specification and differentiation. Nature. 2011; 476(7358):63-68. (Biology). View Reference
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Yu Q, Sharma A, Oh SY, et al. T cell factor 1 initiates the T helper type 2 fate by inducing the transcription factor GATA-3 and repressing interferon-γ. Nat Immunol. 2009; 10(9):992-999. (Biology). View Reference
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