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Multicolor flow cytometric analysis of CD90 expression on rat splenocytes. Lewis rat splenic leucocytes were preincubated with Purified Mouse Anti-Rat CD32 antibody (Rat BD Fc Block™) (Cat. No. 550270/550271). The cells were stained with PE Mouse Anti-Rat CD3 antibody (Cat. No. 554833) and either BD Horizon™ R718 Mouse IgG1, κ Isotype Control (Cat. No. 566928; Left Plot) or BD Horizon™ R718 Mouse Anti-Rat CD90/Mouse CD90.1 antibody (Cat. No. 567361; Right Plot) at 0.5 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of CD90 (or Ig Isotype control staining) versus CD3 was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
BD Horizon™ R718 Mouse Anti-Rat CD90/Mouse CD90.1
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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 CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
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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.
The antibody was conjugated to BD Horizon™ Red 718, which has been developed exclusively by 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 (21)
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