Flow cytometric analysis of the T-cell receptor (TcR) β-chain on mouse splenocytes. Splenocytes from C57BL/6 mice were stained with either a APC-Cy™7 Hamster IgG2, λ1 isotype control (Cat. No. 560604; shaded) or with the APC-Cy™7 Hamster Anti-Mouse TcR β-chain antibody (Cat. No. 560656; unshaded; left panel). Splenocytes from C57BL/6 mice were stained with both a PE Rat Anti-Mouse CD4 antibody (Cat. No. 553049) and a PE Rat Anti-Mouse CD8a antibody (Cat. No. 553033) in conjunction with either a APC-Cy™7 Hamster IgG2, λ1 isotype control (middle panel) or the APC-Cy™7 Hamster Anti-Mouse TcR β-chain antibody (right panel). Histograms and dot plots were derived from gated events based on light scattering characteristics for splenocytes. Flow cytometry was performed on a BD™ LSR II flow cytometry system.
Flow cytometric analysis of the T-cell receptor (TcR) β-chain on mouse splenocytes. Splenocytes from C57BL/6 mice were stained with either a APC-Cy™7 Hamster IgG2, λ1 isotype control (Cat. No. 560604; shaded) or with the APC-Cy™7 Hamster Anti-Mouse TcR β-chain antibody (Cat. No. 560656; unshaded; left panel). Splenocytes from C57BL/6 mice were stained with both a PE Rat Anti-Mouse CD4 antibody (Cat. No. 553049) and a PE Rat Anti-Mouse CD8a antibody (Cat. No. 553033) in conjunction with either a APC-Cy™7 Hamster IgG2, λ1 isotype control (middle panel) or the APC-Cy™7 Hamster Anti-Mouse TcR β-chain antibody (right panel). Histograms and dot plots were derived from gated events based on light scattering characteristics for splenocytes. Flow cytometry was performed on a BD™ LSR II flow cytometry system.
Product Details
BD Pharmingen™
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_1727574
Aqueous buffered solution containing 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 APC-Cy7 under optimum conditions, and unconjugated antibody and free APC-Cy7 were removed.
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.
- Warning: Some APC-Cy7 and PE-Cy7 conjugates show changes in their emission spectrum with prolonged exposure to formaldehyde. If you are unable to analyze fixed samples within four hours, we recommend that you use BD™ Stabilizing Fixative (Cat. No. 338036).
- 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.
- 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.
- APC-Cy7 tandem fluorochrome emission is collected in a detector for fluorescence wavelengths of 750 nm and higher.
- APC-Cy7 is a tandem fluorochrome composed of Allophycocyanin (APC), which is excited by laser lines between 595 and 647 nm and serves as an energy donor, coupled to the cyanine dye Cy7™, which acts as an energy acceptor and fluoresces at 780 nm. BD Biosciences Pharmingen has maximized the fluorochrome energy transfer in APC-Cy7, thus maximizing its fluorescence emission intensity, minimizing residual emission from APC, and minimizing required electronic compensation in multilaser-laser flow cytometry systems. Note: Although every effort is made to minimize the lot-to-lot variation in residual emission from APC, it is strongly recommended that every lot be tested for differences in the amount of compensation required and that individual compensation controls are run for each APC-Cy7 conjugate.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Cy is a trademark of GE Healthcare.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Lysing Solution 10X Concentrate IVD
Size
100 mL
Cat No.
349202
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
APC-Cy™7 Hamster IgG2, λ1 Isotype Control RUO
Size
0.1 mg
Cat No.
560604
PE Rat Anti-Mouse CD4 RUO
Size
0.2 mg
Cat No.
553049
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560656 Rev. 2
Antibody Details
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.
560656 Rev. 2
Format Details
APC-Cy7
APC-Cy7 dye is a part of the BD APC red family of dyes. This tandem fluorochrome is comprised of a Allophycocyanin (APC) donor that has excitation maxima (Ex Max) of 651 nm and an acceptor dye, Cy™7, with an emission maximum (Em Max) at 779 nm. APC-Cy7 can be excited by the Red (627-640 nm) laser and detected using an optical filter centered near 780 nm (e.g., a 760/60 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
APC-Cy7
Red 627-640 nm
651 nm
779 nm
560656 Rev.2
Citations & References
Development References (20)
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Bendelac A, Killeen N, Littman DR, Schwartz RH. A subset of CD4+ thymocytes selected by MHC class I molecules. Science. 1994; 263(5154):1774-1778. (Biology). View Reference
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Bonneville M, Itohara S, Krecko EG, et al. Transgenic mice demonstrate that epithelial homing of gamma/delta T cells is determined by cell lineages independent of T cell receptor specificity. J Exp Med. 1990; 171(4):1015-1026. (Biology). View Reference
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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. (Biology). View Reference
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Davenport C, Kumar V, Bennett M. Rapid rejection of H2k and H2k/b bone marrow cell grafts by CD8+ T cells and NK cells in irradiated mice. J Immunol. 1995; 155(8):3742-3749. (Biology). View Reference
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Duke RC, Cohen JJ, Boehme SA, et al. Morphological, biochemical, and flow cytometric assays of apoptosis. In: Coligan J, Kruisbeek AM, Margulies D, Shevach EM, Strober W, ed. Current Protocols in Immunology. New York: John Wiley and Sons; 1995:3.17.1-3.17.33.
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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. (Biology: 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. (Methodology: 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: Flow cytometry). View Reference
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Lefrancois L. Phenotypic complexity of intraepithelial lymphocytes of the small intestine. J Immunol. 1991; 147(6):1746-1751. (Biology). View Reference
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Ohno H, Ono S, Hirayama N, Shimada S, Saito T. Preferential usage of the Fc receptor gamma chain in the T cell antigen receptor complex by gamma/delta T cells localized in epithelia. J Exp Med. 1994; 179(1):365-369. (Biology). 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. (Biology). View Reference
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Saint-Ruf C, Ungewiss K, Groettrup M, Bruno L, Fehling HJ, von Boehmer H. Analysis and expression of a cloned pre-T cell receptor gene. Science. 1994; 266(5188):1208-1212. (Biology: Stimulation). View Reference
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Skeen MJ, Ziegler HK. Induction of murine peritoneal gamma/delta T cells and their role in resistance to bacterial infection. J Exp Med. 1993; 178(3):971-984. (Biology). View Reference
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Takizawa F, Kinet JP, Adamczewski M. Binding of phycoerythrin and its conjugates to murine low affinity receptors for immunoglobulin G. J Immunol Methods. 1993; 162(2):269-272. (Biology). View Reference
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Vicari AP, Zlotnik A. Mouse NK1.1+ T cells: a new family of T cells. Immunol Today. 1996; 17(2):71-76. (Biology). View Reference
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Wagner DH Jr, Hagman J, Linsley PS, Hodsdon W, Freed JH, Newell MK. Rescue of thymocytes from glucocorticoid-induced cell death mediated by CD28/CTLA-4 costimulatory interactions with B7-1/B7-2. J Exp Med. 1996; 184(5):1631-1638. (Biology). View Reference
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van Vugt MJ, van den Herik-Oudijk IE, van de Winkle JG. Binding of PE-CY5 conjugates to the human high-affinity receptor for IgG (CD64). Blood. 1996; 88(6):2358-2361. (Biology). View Reference
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van der Heyde HC, Elloso MM, Chang WL, Kaplan M, Manning DD, Weidanz WP. Gamma delta T cells function in cell-mediated immunity to acute blood-stage Plasmodium chabaudi adami malaria. J Immunol. 1995; 154(8):3985-3990. (Biology). View Reference
560656 Rev. 2
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.
