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Multicolor flow cytometric analysis of CD4 expression on rat splenocytes. Splenocytes from a Lewis rat were stained with the PE-Cy™7 Mouse Anti-Rat CD4 antibody (Cat. No. 561578) in conjunction with a PE Mouse Anti-Rat CD3 antibody (Cat. No. 554833). The two-color flow cytometric dot plot shows CD3 versus CD4 expression by cells with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry was performed on a BD LSRII™ System.
BD Pharmingen™ PE-Cy™7 Mouse Anti-Rat CD4
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
PE-Cy™7 tandem fluorochrome emission is collected in a detector for fluorescence wavelengths of 750 nm and higher. PE-Cy™7-labeled antibodies can be used with FITC- and R-PE-labeled reagents in single-laser flow cytometers with no significant spectral overlap between PE-Cy™7 and FITC.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Cy is a trademark of Amersham Biosciences Limited. This conjugated product is sold under license to the following patents: US Patent Nos. 5,486,616; 5,569,587; 5,569,766; 5,627,027.
- 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.
- 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.
- 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).
- This product is subject to proprietary rights of Amersham Biosciences Corp. and Carnegie Mellon University and made and sold under license from Amersham Biosciences Corp. This product is licensed for sale only for research. It is not licensed for any other use. If you require a commercial license to use this product and do not have one return this material, unopened to BD Biosciences, 10975 Torreyana Rd, San Diego, CA 92121 and any money paid for the material will be refunded.
- PE-Cy7 is a tandem fluorochrome composed of R-phycoerythrin (PE), which is excited by 488-nm light and serves as an energy donor, coupled to the cyanine dye Cy7, which acts as an energy acceptor and fluoresces maximally at 780 nm. PE-Cy7 tandem fluorochrome emission is collected in a detector for fluorescence wavelengths of 750 nm and higher. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from PE may be observed. Therefore, we recommend that individual compensation controls be performed for every PE-Cy7 conjugate. PE-Cy7 is optimized for use with a single argon ion laser emitting 488-nm light, and there is no significant overlap between PE-Cy7 and FITC emission spectra. When using dual-laser cytometers, which may directly excite both PE and Cy7, we recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
Companion Products
The OX-35 clone recognizes the CD4 antigen on most thymocytes, a subpopulation of mature T lymphocytes (i.e., MHC class II-restricted T cells, including most T helper cells), monocytes, macrophages, some dendritic cells, and microglia. CD4 is an antigen coreceptor on the T-cell surface that interacts with MHC class II molecules on antigen-presenting cells. It participates in T-cell activation through it's association with the T-cell receptor complex and protein tyrosine kinase Lck. The OX-35 clone has been reported to bind to a different epitope of CD4 than that recognized by the W3/25 and OX-38 clones.
Development References (7)
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Bañuls MP, Alvarez A, Ferrero I, Zapata A, Ardavin C. Cell-surface marker analysis of rat thymic dendritic cells. Immunology. 1993; 79(2):298-304. (Biology). View Reference
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Bierer BE, Sleckman BP, Ratnofsky SE, Burakoff SJ. The biologic roles of CD2, CD4, and CD8 in T-cell activation. Annu Rev Immunol. 1989; 7:579-599. (Biology). View Reference
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Ford AL, Foulcher E, Goodsall AL, Sedgwick JD. Tissue digestion with dispase substantially reduces lymphocyte and macrophage cell-surface antigen expression. J Immunol Methods. 1996; 194(1):71-75. (Biology). View Reference
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Janeway CA Jr. The T cell receptor as a multicomponent signalling machine: CD4/CD8 coreceptors and CD45 in T cell activation. Annu Rev Immunol. 1992; 10:645-674. (Biology). View Reference
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Jefferies WA, Green JR, Williams AF. Authentic T helper CD4 (W3/25) antigen on rat peritoneal macrophages. J Exp Med. 1985; 162(1):117-127. (Immunogen: Flow cytometry, Functional assay, Immunoaffinity chromatography, Immunoprecipitation, Inhibition). View Reference
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Liu L, Zhang M, Jenkins C, MacPherson GG. Dendritic cell heterogeneity in vivo: two functionally different dendritic cell populations in rat intestinal lymph can be distinguished by CD4 expression. J Immunol. 1998; 161(3):1146-1155. (Biology). View Reference
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Wang CC, Wu CH, Shieh JY, Wen CY, Ling EA. Immunohistochemical study of amoeboid microglial cells in fetal rat brain. J Anat. 1996; 189(3):567-574. (Biology). View Reference
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.