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Multiparameter flow cytometric analysis of CD45 expression on Rhesus macaque peripheral blood leukocytes. Rhesus macaque whole blood was stained with a BD Horizon™ V500 Mouse IgG1, κ Isotype Control (Cat. No. 560787; Left Panel) or with BD Horizon™ V500 Mouse Anti-NHP CD45 antibody (Cat. No. 561489; Right Panel). The erythrocytes were lysed with BD PharmLyse™ Lysing Buffer (Cat. No. 555899). Two-parameter flow cytometric dot blots showing the correlated expression of CD45 (or Ig isotype control staining) versus side scattered-light characteristics were derived from events gated for all viable leukocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
BD Horizon™ V500 Mouse Anti-NHP CD45
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
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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- BD Horizon V500 has a maximum absorption of 415 nm and maximum emission of 500 nm. Before staining with this reagent, please confirm that your flow cytometer is capable of exciting the fluorochrome and discriminating the resulting fluorescence.
- BD Horizon V500 is covered by one or more of the following US patents: 8,431,416.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
D058-1283 is a CD45 monoclonal antibody specific for non-human primate leucocytes. It was developed using Rhesus peripheral whole blood as the immunogen. It does not cross-react with human leucocytes. This antibody reacts with baboon, Rhesus and Cynomolgus Macaque leucocytes in a similar pattern to CD45 binding to leukocyte common antigen (LCA) on human cells. Immunophenotypic analysis shows that D058-1283 binds to lymphocytes, monocytes and granulocytes of non-human primate blood samples. This antibody is able to block the binding of monoclonal antibody TÜ116; a reported anti-human CD45 antibody that cross-reacts with nonhuman primate leucocytes. In Western blot analysis, the D058-1283 antibody identifies a 180-200 kDa band.
The antibody is conjugated to BD Horizon™ V500, which has been developed for use in multicolor flow cytometry experiments and is
available exclusively from BD Biosciences. It is excited by the Violet laser with an Ex max of 415 nm and Em Max at 500 nm. BD Horizon V500 conjugates emit at a similar wavelength to Amcyan yet exhibit reduced spillover into the FITC channel. For more information on BD Horizon V500, visit bdbiosciences.com/colors.
When compensating dyes in this spectral range (such as Horizon™ V500 and AmCyan), the most accurate compensation can be obtained using single stained cellular controls. Due to spectral differences between cells and beads in this channel, using BD CompBeads can result in spillover errors for V500 and AmCyan reagents. Therefore, the use of BD CompBeads or BD CompBeads Plus to determine spillover values for these reagents is not recommended. Different V500 reagents (e.g. CD4 vs. CD45) can have slightly different fluorescence spillover therefore, it may also be necessary to use clone specific compensation controls when using these reagents.
Development References (3)
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Kishihara K, Penninger J, Wallace VA, et al. Normal B lymphocyte development but impaired T cell maturation in CD45-exon6 protein tyrosine phosphatase-deficient mice. Cell. 1993; 74(1):143-156. (Biology). View Reference
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Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
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Reimann KA, Waite BC, Lee-Parritz DE, et al. Use of human leukocyte-specific monoclonal antibodies for clinically immunophenotyping lymphocytes of rhesus monkeys. Cytometry. 1994; 17(1):102-108. (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.