Multicolor flow cytometric analysis of CD90 expression on rat splenocytes. Splenocytes from a Lewis rat were stained with the PE-Cy™7 Mouse Anti-Rat CD90 antibody (Cat. No. 561404) in conjunction with a FITC Rat Anti-Mouse CD3 antibody (Cat. No. 554832/559975). A two-color flow cytometric dot plot showing the correlated expression of CD90 versus CD3 was derived from gated events based on the forward and side light-scatter characteristics of viable splenocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometry System.
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Multicolor flow cytometric analysis of CD90 expression on rat splenocytes. Splenocytes from a Lewis rat were stained with the PE-Cy™7 Mouse Anti-Rat CD90 antibody (Cat. No. 561404) in conjunction with a FITC Rat Anti-Mouse CD3 antibody (Cat. No. 554832/559975). A two-color flow cytometric dot plot showing the correlated expression of CD90 versus CD3 was derived from gated events based on the forward and side light-scatter characteristics of viable splenocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometry System.
Product Details
BD Pharmingen™
Rat Thy-1; Mouse Thy-1.1
Rat (QC Testing), Mouse (Tested in Development)
Mouse BALB/c IgG1, κ
Rat Thymocyte Thy-1 Antigen
Flow cytometry (Routinely Tested)
0.2 mg/ml
24832,21838
AB_10645378
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with PE-Cy7 under optimum conditions, and unconjugated antibody and free PE-Cy7 were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Recommended Assay Procedures
PE-Cy7 tandem fluorochrome emission is collected in a detector for fluorescence wavelengths of 750 nm and higher. PE-Cy7-labeled antibodies can be used with FITC- and R-PE-labeled reagents in single-laser flow cytometers with no significant spectral overlap between PE-Cy7 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
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
FITC Mouse Anti-Rat CD3 RUO
Size
0.5 mg
Cat No.
554832
FITC Mouse Anti-Rat CD3 RUO
Size
0.1 mg
Cat No.
559975
561404 Rev. 1
Antibody Details
OX-7
CD90 (Thy-1) is a GPI-anchored membrane glycoprotein of the Ig superfamily which is involved in signal transduction. The OX-7 monoclonal antibody specifically binds to rat CD90 reported to be expressed by hematopoietic stem cells, early myeloid and erythroid cells, immature B lymphocytes in the bone marrow and peripheral lymphoid organs, thymocytes, recent thymic emigrants (a subset of CD45RC- peripheral T lymphocytes), neurons, glomerular mesangial cells, endothelium at inflammatory sites, mast cells, and dendritic cells. Rat dendritic epidermal T cells (DEC) have been reported to be CD90 (Thy-1) negative, unlike those of the mouse.
The OX-7 clone has been reported to crossreact with the mouse CD90.1 (Thy-1.1) alloantigen of the AKR/J and PL strains, but not CD90.2 (Thy-1.2) found on many mouse strains. In the mouse, CD90 is found on thymocytes, most peripheral T lymphocytes, some intraepithelial T lymphocytes (IEL, DEC), hematopoietic stem cells, and neurons, but not B lymphocytes. In addition, there is evidence that CD90 mediates adhesion of mouse thymocytes to mouse thymic stroma. The OX-7 clone has also been reported to crossreact with rabbit and guinea pig thymus, brain, and intestine.
561404 Rev. 1
Format Details
PE-Cy7
PE-Cy7 dye is a part of the BD PE family of dyes. This tandem fluorochrome is comprised of a R-Phycoerythrin (PE) donor that has excitation maxima (Ex Max) of 496-nm and 566-nm and an acceptor dye, Cy™7, with an emission maximum (Em Max) at 781-nm. PE can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and detected using an optical filter centered near 781 nm (e.g., a 760/60-nm bandpass filter). The donor dye can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and the acceptor dye can be excited by the Red (627–640-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
PE-Cy7
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
781 nm
561404 Rev.1
Citations & References
Development References (18)
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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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Campbell DG, Gagnon J, Reid KB, Williams AF. Rat brain Thy-1 glycoprotein. The amino acid sequence, disulphide bonds and an unusual hydrophobic region. Biochem J. 1981; 195(1):15-30. (Biology). View Reference
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Chen-Woan M, Delaney CP, Fournier V, et al. In vitro characterization of rat bone marrow-derived dendritic cells and their precursors. J Leukoc Biol. 1996; 59(2):196-207. (Biology). View Reference
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Crook K and Hunt SV. Enrichment of early fetal-liver hemopoietic stem cells of the rat using monoclonal antibodies against the transferrin receptor, Thy-1, and MRC-OX82. Dev Immunol. 1996; 4:235-246. (Biology). View Reference
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Dráberová L, Amoui M, and Dráber P. Thy-1-mediated activation of rat mast cells: the role of Thy-1 membrane microdomains. Immunology. 1996; 87(1):141-148. (Clone-specific: Activation, Western blot). View Reference
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Elbe A, Kilgus O, Hünig T, and Stingl G. T-cell receptor diversity in dendritic epidermal T cells in the rat. J Invest Dermatol. 1993; 102:74-79. (Biology). View Reference
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Ferrero I, Bañnuls M, Alvarez A, Ardavín C. Rat thymic dendritic cells: cell surface marker variations in culture. Immunol Lett. 1993; 37(2-3):241-247. (Biology). View Reference
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Garnett D, Barclay AN, Carmo AM, Beyers AD. The association of the protein tyrosine kinases p56lck and p60fyn with the glycosyl phosphatidylinositol-anchored proteins Thy-1 and CD48 in rat thymocytes is dependent on the state of cellular activation. Eur J Immunol. 1993; 23(10):2540-2544. (Clone-specific: Immunoprecipitation). View Reference
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He HT, Naquet P, Caillol D, Pierres M. Thy-1 supports adhesion of mouse thymocytes to thymic epithelial cells through a Ca2(+)-independent mechanism. J Exp Med. 1991; 173(2):515-518. (Biology). View Reference
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Hermans MH, Opstelten D. In situ visualization of hemopoietic cell subsets and stromal elements in rat and mouse bone marrow by immunostaining of frozen sections. J Histochem Cytochem. 1991; 39(12):1627-1634. (Clone-specific: Immunohistochemistry). View Reference
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Hosseinzadeh H, Goldschneider I. Recent thymic emigrants in the rat express a unique antigenic phenotype and undergo post-thymic maturation in peripheral lymphoid tissues. J Immunol. 1993; 150(5):1670-1679. (Biology). View Reference
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Ishizu A, Ishikura H, Nakamaru Y et al. Thy-1 induced on rat endothelium regulates vascular permeability at sites of inflammation. Int Immunol. 1995; 7:1939-1947. (Clone-specific: Immunoprecipitation). View Reference
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Kawachi H, Orikasa M, Matsui K, et al. Epitope-specific induction of mesangial lesions with proteinuria by a MoAb against mesangial cell surface antigen. Clin Exp Immunol. 1992; 88(3):399-404. (Clone-specific: Western blot). 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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Mason DW, Williams AF. The kinetics of antibody binding to membrane antigens in solution and at the cell surface. Biochem J. 1980; 187(1):1-20. (Immunogen). View Reference
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Nakashima I, Zhang YH, Rahman SM, et al. Evidence of synergy between Thy-1 and CD3/TCR complex in signal delivery to murine thymocytes for cell death. J Immunol. 1991; 147(4):1153-1162. (Biology). View Reference
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Paul LC, Rennke HG, Milford EL, and Carpenter CB. Thy-1.1 in glomeruli of rat kidneys. Kidney Int. 1984; 25:771-777. (Clone-specific: Electron microscopy, Immunohistochemistry). View Reference
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Payer E, Elbe A, Stingl G. Circulating CD3+/T cell receptor V gamma 3+ fetal murine thymocytes home to the skin and give rise to proliferating dendritic epidermal T cells. J Immunol. 1991; 146(8):2536-2543. (Biology). View Reference
561404 Rev. 1
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.
