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Flow cytometric analysis of CD45 expression on mouse splenocytes. Splenic leucocytes from BALB/c mice were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were stained with either BD Horizon™ BV510 Rat IgG2b, κ Isotype Control (Cat No. 562951; dashed line histogram) or BD Horizon™ BV510 Rat Anti-Mouse CD45 antibody (Cat No. 567800; solid line histogram) at 0.5 µg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The fluorescence histogram showing CD45 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) mouse 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™ BV510 Rat Anti-Mouse 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
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.
For optimal and reproducible results, BD Horizon Brilliant™ Stain Buffer should be used anytime BD Horizon Brilliant™ dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant™ Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant™ Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant™ Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant™ Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant™ Stain Buffer Plus (Cat. No. 566385).
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
- BD Horizon Brilliant Violet 510 is covered by one or more of the following US patents: 8,575,303; 8,354,239.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
Companion Products
The mAb I3/2.3 recognizes mouse CD45, which is also known as the leukocyte common antigen (L-CA or LCA), T200, or Lymphocyte antigen 5 (Ly-5). CD45 is a 180-240 kDa single-pass type I transmembrane glycoprotein that is encoded by Ptprc (protein tyrosine phosphatase, receptor type, C) which belongs to the protein tyrosine phosphatase (PTP) family. CD45 is variably expressed on thymocytes and other hematopoietic cells except mature erythrocytes and platelets. Different isoforms of CD45 arise from alternative splicing of variable exons 4, 5, and 6, which encode A, B, and C determinants, respectively, that encode extracellular domains of the transmembrane receptor. The differential expression of these isoforms is related to the cell's lineage as well as its state of activation and maturation. The mAb I3/2.3 reportedly recognizes all CD45 isoforms. CD45 plays key roles as a protein tyrosine-protein phosphatase in leucocyte responses including signal pathways transduced by antigen specific receptors expressed on T cells (TCRs) and B cells (BCRs).
The antibody was conjugated to BD Horizon™ BV510 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 405-nm and Em Max at 510-nm, BD Horizon™ BV510 can be excited by the violet laser and detected in the BD Horizon™ V500 (525/50-nm) filter set. BD Horizon™ BV510 conjugates are useful for the detection of dim markers off the violet laser.
Development References (5)
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Alaverdi N. Monoclonal antibodies to mouse cell-surface antigens.. Curr Protoc Immunol. 2002; Appendix 4:Appendix 4B. (Clone-specific). View Reference
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Johnson P, Greenbaum L, Bottomly K, Trowbridge IS. Identification of the alternatively spliced exons of murine CD45 (T200) required for reactivity with B220 and other T200-restricted antibodies. J Exp Med. 1989; 169(3):1179-1184. (Clone-specific: Flow cytometry). View Reference
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Lefrancois L, Goodman T. Developmental sequence of T200 antigen modifications in murine T cells.. J Immunol. 1987; 139(11):3718-24. (Clone-specific: Flow cytometry, Immunoprecipitation, Radioimmunoassay). View Reference
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Takedachi M, Qu D, Ebisuno Y, et al. CD73-generated adenosine restricts lymphocyte migration into draining lymph nodes.. J Immunol. 2008; 180(9):6288-96. (Biology). View Reference
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Trowbridge IS. Interspecies spleen-myeloma hybrid producing monoclonal antibodies against mouse lymphocyte surface glycoprotein, T200.. J Exp Med. 1978; 148(1):313-23. (Immunogen: Immunoprecipitation, Ribonuclease protection assay). View Reference
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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.