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Flow cytometric analysis of CD3 expression on Rhesus monkey peripheral blood leucocytes. Rhesus whole blood was stained with either BD Horizon™ R718 Mouse IgG1, κ Isotype Control (Cat. No. 566928; Left Plot) or BD Horizon™ R718 Mouse Anti-Human/NHP CD3 antibody (Cat. No. 566955/566956; Right Plot). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). A two-parameter pseudocolor density plot showing the correlated expression of CD3 (or Ig Isotype control staining) versus side light-scatter (SSC) signals was derived from gated events with the forward and side light-scatter characteristics of intact leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Flow Cytometer System and FlowJo™ software.
BD Horizon™ R718 Mouse Anti-Human CD3
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 CompBead to ensure that BD CompBeads are appropriate for your specific cellular application.
Product Notices
- 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).
- 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.
- Alexa Fluor® is a registered trademark of Life Technologies Corporation.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- This product is provided under an Agreement between BIOTIUM and BD Biosciences. 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.
Companion Products
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.
The antibody was conjugated to BD Horizon Red 718, which has been developed exclusively for BD Biosciences as a better alternative to Alexa Fluor® 700. BD Horizon Red 718 can be excited by the red laser (628 – 640 nm) and, with an Em Max around 718 nm, it can be detected using a 730/45 nm filter. Due to similar excitation and emission properties, we do not recommend using R718 in combination with APC-R700 or Alexa Fluor® 700.
Development References (7)
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Bleavins MR, Brott DA, Alvey JD, de la Iglesia FA. Flow cytometric characterization of lymphocyte subpopulations in the cynomolgus monkey (Macaca fascicularis). Vet Immunol Immunopathol. 1993; 37(1):1-13. (Biology). View Reference
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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: Immunoprecipitation, Western blot). View Reference
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Conrad ML, Davis WC, Koop BF. TCR and CD3 antibody cross-reactivity in 44 species. Cytometry A. 2007; 71(11):925-933. (Biology). View Reference
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Engel P, Wagner N, Tedder TF. CD86 Workshop Report. In: 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; 1995:703-705.
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Jacobsen CN, Aasted B, Broe MK, Petersen JL. Reactivities of 20 anti-human monoclonal antibodies with leucocytes from ten different animal species. Vet Immunol Immunopathol. 1993; 39(4):461-466. (Biology). View Reference
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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: Activation, Dot Blot, Functional assay, Immunoprecipitation, Western blot). View Reference
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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.
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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.