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APC-Cy™7 Mouse Anti-Human CD3
APC-Cy™7 Mouse Anti-Human CD3

Flow cytometric analysis of CD3 expression on Rhesus macaque (Macaca mulatta) peripheral blood lymphocytes. Whole rhesus blood was stained with either APC-Cy™7 Mouse IgG1, κ Isotype Control (Cat. No. 557873; dashed line histogram) or APC-Cy™7 Mouse Anti-Human CD3 (Cat. No. 557757; solid line histogram). Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). Fluorescent histograms were derived from gated events with the side and forward light-scatter characteristics of viable lymphocytes.

Flow cytometric analysis of CD3 expression on Rhesus macaque (Macaca mulatta) peripheral blood lymphocytes. Whole rhesus blood was stained with either APC-Cy™7 Mouse IgG1, κ Isotype Control (Cat. No. 557873; dashed line histogram) or APC-Cy™7 Mouse Anti-Human CD3 (Cat. No. 557757; solid line histogram). Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). Fluorescent histograms were derived from gated events with the side and forward light-scatter characteristics of viable lymphocytes.

Product Details
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BD Pharmingen™
CD3E; CD3-epsilon; T3E; TCRE
Rhesus, Cynomolgus, Baboon (QC Testing), Human (Tested in Development)
Mouse BALB/c IgG1, λ
Purified Human CD3ε Protein
Flow cytometry (Routinely Tested), Induction (Reported)
5 µl
AB_396863
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 APC-Cy7 under optimum conditions, and unconjugated antibody and free APC-Cy7 were removed.

Product Notices

  1. 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).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. 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.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. 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.
  7. APC-Cy7 tandem fluorochrome emission is collected in a detector for fluorescence wavelengths of 750 nm and higher.
  8. 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.
  9. 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).
  10. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  11. Cy is a trademark of GE Healthcare.
  12. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
557757 Rev. 8
Antibody Details
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SP34-2

Clone SP34-2 is a mouse IgG1 isotype monoclonal antibody, descendant of SP34 (mouse IgG3), with the same specificity and reactivity pattern as the parent clone. It cross-reacts with a major subset of peripheral blood lymphocytes, but not monocytes or granulocytes, of baboon, and rhesus, cynomolgus, and pigtail macaque monkeys. The distribution on lymphocytes is similar to that observed with normal human donor lymphocytes with the majority of CD3-positive cells being negative when dual stained with antibodies to B or NK cells markers. SP34-2 is also capable of inducing cell proliferation on both human and non-human primate PBMC.

557757 Rev. 8
Format Details
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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.
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APC-Cy7
Red 627-640 nm
651 nm
779 nm
557757 Rev.8
Citations & References
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Development References (8)

  1. Beavis AJ, Pennline KJ. Allo-7: a new fluorescent tandem dye for use in flow cytometry. Cytometry. 1996; 24(4):390-395. (Biology). View Reference
  2. Blumberg RS, Ley S, Sancho J, et al. Structure of the T-cell antigen receptor: evidence for two CD3 epsilon subunits in the T-cell receptor-CD3 complex. Proc Natl Acad Sci U S A. 1990; 87(18):7220-7224. (Clone-specific). View Reference
  3. Carter DL, Shieh TM, Blosser RL et al. CD56 identifies monocytes and not natural killer cells in rhesus macaques. Cytometry. 1999; 37(1):41-50. (Biology). View Reference
  4. Pessano S, Oettgen H, Bhan AK, Terhorst C. The T3/T cell receptor complex: antigenic distinction between the two 20-kd T3 (T3-delta and T3-epsilon) subunits. EMBO J. 1985; 4(2):337-344. (Immunogen). View Reference
  5. Roederer M, Kantor AB, Parks DR, Herzenberg LA. Cy7PE and Cy7APC: bright new probes for immunofluorescence. Cytometry. 1996; 24(3):191-197. (Biology). View Reference
  6. Sancho J, Ledbetter JA, Choi MS, Kanner SB, Deans JP, Terhorst C. CD3-zeta surface expression is required for CD4-p56lck-mediated upregulation of T cell antigen receptor-CD3 signaling in T cells. J Biol Chem. 1992; 267(11):7871-7879. (Biology). View Reference
  7. Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995.
  8. Wilson AD, Shooshtari M, Finerty S, Watkins P, Morgan AJ. Selection of monoclonal antibodies for the identification of lymphocyte surface antigens in the New World primate Saguinus oedipus oedipus (cotton top tamarin). J Immunol Methods. 1995; 178(2):195-200. (Clone-specific). View Reference
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557757 Rev. 8

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.