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PE-CF594 Hamster Anti-Mouse TCR β Chain
PE-CF594 Hamster Anti-Mouse TCR β Chain
Two-color flow cytometric analysis of TCRβ expression on mouse splenocytes. Mouse splenic leucocytes were incubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with APC Rat Anti-Mouse CD4 (Cat. No. 553051/561091) and APC Rat Anti-Mouse CD8a (Cat. No. 553035/561093) antibodies and either BD Horizon™ PE-CF594 Hamster IgG2, λ1 Isotype Control (Cat. No. 562522, Left Panel) or BD Horizon™ PE-CF594 Hamster Anti-Mouse TCR β Chain antibody (Cat. No. 562841; Right Panel). Two-color flow cytometric dot plots show the correlated expression patterns of TCRβ (or Ig Isotype control staining) versus CD4 and CD8a for gated events with the forward and side light-scatter characteristics of viable splenic leucocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Two-color flow cytometric analysis of TCRβ expression on mouse splenocytes. Mouse splenic leucocytes were incubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with APC Rat Anti-Mouse CD4 (Cat. No. 553051/561091) and APC Rat Anti-Mouse CD8a (Cat. No. 553035/561093) antibodies and either BD Horizon™ PE-CF594 Hamster IgG2, λ1 Isotype Control (Cat. No. 562522, Left Panel) or BD Horizon™ PE-CF594 Hamster Anti-Mouse TCR β Chain antibody (Cat. No. 562841; Right Panel). Two-color flow cytometric dot plots show the correlated expression patterns of TCRβ (or Ig Isotype control staining) versus CD4 and CD8a for gated events with the forward and side light-scatter characteristics of viable splenic leucocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Product Details
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BD Horizon™
Tcrb; TCRbeta; TCRβ, T cell receptor beta chain
Mouse (QC Testing)
Armenian Hamster IgG2, λ1
TCR affinity-purified from mouse T-cell hybridoma DO-11.10
Flow cytometry (Routinely Tested)
0.2 mg/ml
21577
AB_2737831
Aqueous buffered solution containing BSA, protein stabilizer, and ≤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 with BD Horizon™ PE-CF594 under optimum conditions, and unconjugated antibody and free PE-CF594 were removed.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  3. An isotype control should be used at the same concentration as the antibody of interest.
  4. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  5. 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.
  6. 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.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. Texas Red is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  9. CF™ is a trademark of Biotium, Inc.
  10. When excited by the yellow-green (561-nm) laser, the fluorescence may be brighter than when excited by the blue (488-nm) laser.
  11. This product is provided under an Agreement between BIOTIUM and BD Biosciences. The manufacture, use, sale, offer for sale, or import of this product is subject to one or more patents or pending applications owned or licensed by Biotium, Inc. 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.
  12. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using multi-laser cytometers, which may directly excite both PE and CF™594.
  13. Although hamster immunoglobulin isotypes have not been well defined, BD Biosciences Pharmingen has grouped Armenian and Syrian hamster IgG monoclonal antibodies according to their reactivity with a panel of mouse anti-hamster IgG mAbs. A table of the hamster IgG groups, Reactivity of Mouse Anti-Hamster Ig mAbs, may be viewed at http://www.bdbiosciences.com/documents/hamster_chart_11x17.pdf.
562841 Rev. 1
Antibody Details
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H57-597

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.

This antibody is conjugated to BD Horizon™ PE-CF594, which has been developed exclusively by BD Biosciences as a better alternative to PE-Texas Red®. PE-CF594 excites and emits at similar wavelengths to PE-Texas Red® yet exhibits improved brightness and spectral characteristics. Due to PE having maximal absorption peaks at 496 nm and 564 nm, PE-CF594 can be excited by the blue (488-nm), green (532-nm) and yellow-green (561-nm) lasers and can be detected with the same filter set as PE-Texas Red® (eg 610/20-nm filter).

562841 Rev. 1
Format Details
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PE-CF594
BD Horizon™ PE-CF594 dye is a part of the BD PE family of dyes. This tandem fluorochrome is comprised of a R-Phycoerythrin (PE) donor that has excitation maxima (Ex Max) of 496-nm and 566-nm and an acceptor dye with an emission maximum (Em Max) at 615-nm. PE-CF594, driven by BD innovation, is designed to be excited by the blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and detected using an optical filter centered near 615 nm (e.g., a 610/20-nm bandpass filter). The donor dye can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and the acceptor dye can be excited by the green (532-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PE-CF594
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
615 nm
562841 Rev.1
Citations & References
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Development References (5)

  1. 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
  2. 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
  3. 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
  4. 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
  5. 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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562841 Rev. 1

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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.