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Two-color flow cytometric analysis of TCRβ expression on mouse splenocytes. Mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with PE Rat Anti-Mouse CD4 (Cat. No. 553048/553049/561837) and PE Rat Anti-Mouse CD8a (Cat. No. 553033/553032/561095) antibodies and either BD Horizon™ BUV737 Hamster IgG2, λ1 Isotype Control (Cat. No. 612822; Left Plot) or BD Horizon BUV737 Hamster Anti-Mouse TCR β Chain antibody (Cat. No. 612821; Right Plot) at 0.5 µg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The two-color flow cytometric dot plot showing the correlated expression of TCRβ (or Ig Isotype control staining) versus CD4 and CD8a was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) splenic leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
BD Horizon™ BUV737 Hamster Anti-Mouse TCR β Chain
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. This will 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 two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. 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).
Note: When using high concentrations of antibody, background binding of this dye to erythroid cell subsets (mature erythrocytes and precursors) has been observed. For researchers studying these cell populations, or in cases where light scatter gating does not adequately exclude these cells from the analysis, this background may be an important factor to consider when selecting reagents for panel(s).
Product Notices
- 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.
- 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.
- BD Horizon Brilliant Ultraviolet 737 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,575,303; 8,354,239.
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
The H57-597 antibody reacts with a common epitope of the β chain of the T-cell Receptor (TCR) complex on αβ TCR-expressing thymocytes, peripheral T lymphocytes, NK1.1+ thymocytes, and NK-T cells of all mouse strains tested. It does not react with γδ TCR-bearing T cells. In the fetal and adult thymus, the TCR β-chain may form homodimers or pair with the pre-TCR α-chain on the surface of immature thymocytes before TCR α-chain expression. Plate-bound or soluble H57-597 antibody activates αβ TCR-bearing T cells, and plate-bound mAb can induce apoptotic death.
The antibody was conjugated to BD Horizon BUV737 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome with an Ex Max near 350 nm and an Em Max near 737 nm. BD Horizon Brilliant BUV737 can be excited by the ultraviolet laser (355 nm) and detected with a 740/35 nm filter. Due to the excitation of the acceptor dye by the red laser line, there may be significant spillover into red laser detectors with filters in the 700-720 nm range.
Development References (5)
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Castro JE, Listman JA, Jacobson BA, et al. Fas modulation of apoptosis during negative selection of thymocytes. Immunity. 1996; 5(6):617-627. (Clone-specific: Flow cytometry, Fluorescence microscopy, Immunofluorescence). View Reference
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Gascoigne NR. Transport and secretion of truncated T cell receptor beta-chain occurs in the absence of association with CD3. J Biol Chem. 1990; 265(16):9296-9301. (Clone-specific: Immunofluorescence, Immunoprecipitation). View Reference
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Kruisbeek AM, Shevach EM. Proliferative assays for T cell function. Curr Protoc Immunol. 2004; 3:3.12.1-3.12.14. (Clone-specific: Activation, Functional assay, Stimulation). View Reference
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Kubo RT, Born W, Kappler JW, Marrack P, Pigeon M. Characterization of a monoclonal antibody which detects all murine alpha beta T cell receptors. J Immunol. 1989; 142(8):2736-2742. (Immunogen: Activation, Flow cytometry, Functional assay, Stimulation). View Reference
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Saint-Ruf C, Panigada M, Azogui O, Debey P, von Boehmer H, Grassi F. Different initiation of pre-TCR and gammadeltaTCR signalling. Nature. 2000; 406(6795):524-527. (Clone-specific: Flow cytometry, Fluorescence microscopy, Immunofluorescence, Immunoprecipitation, Western blot). 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.