Flow cytometric analysis of CD3 expression on Rhesus macaque (Macaca mulatta) peripheral blood lymphoyctes. Rhesus whole blood was stained with either Pacific Blue™ Mouse Anti-Human CD3 (Cat. No.558124; solid line histogram) or Pacific Blue™ Mouse IgG1 κ Isotype Control (Cat. No. 558120; dashed line histogram). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Fluorescent histograms were derived from gated events with the side and forward light-scattering characteristics of viable lymphocytes.
Flow cytometric analysis of CD3 expression on Rhesus macaque (Macaca mulatta) peripheral blood lymphoyctes. Rhesus whole blood was stained with either Pacific Blue™ Mouse Anti-Human CD3 (Cat. No.558124; solid line histogram) or Pacific Blue™ Mouse IgG1 κ Isotype Control (Cat. No. 558120; dashed line histogram). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Fluorescent histograms were derived from gated events with the side and forward light-scattering characteristics of viable lymphocytes.
Product Details
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)
0.2 mg/ml
AB_397044
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 is conjugated to Pacific Blue™ under optimum conditions, and unreacted Pacific Blue™ was 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
- Pacific Blue™ has a maximum absorption of 416 nm and maximum emission of 451 nm. Before staining with this reagent, please confirm that your flow cytometer is capable of exciting the fluorochrome and discriminating the resulting fluorescence.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
- 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 Buffer RUO
Size
100 mL
Cat No.
555899
Pacific Blue™ Mouse IgG1, κ Isotype Control RUO
Size
0.1 mg
Cat No.
558120
558124 Rev. 8
Antibody Details
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.
558124 Rev. 8
Format Details
Pacific Blue
Pacific Blue™ is part of the BD violet family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 404-nm and an emission maximum (Em Max) at 455-nm. Pacific Blue is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 450-nm (e.g., a 450/50-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Pacific Blue
Violet 405 nm
404 nm
455 nm
558124 Rev.8
Citations & References
Development References (6)
-
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
-
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
-
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
-
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
-
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.
-
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. (Biology). View Reference
558124 Rev. 8
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.
