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Flow cytometric analysis of CD135 expression on mouse bone-marrow leukocytes. BALB/c bone marrow cells were costained with FITC Mouse Anti-Rat CD11b (Cat. No. 554982) and FITC Rat Anti-Mouse CD45R/B220 (Cat. No. 553087), and either PE Rat IgG2a, κ Isotype Control (Cat. No. 553930; left panel) or PE Rat Anti-Mouse CD135 (Cat. No. 561068/553842; right panel). Fluorescent dot plots were derived from gated events with the side and forward light-scattering characteristics of viable leukocytes. Flow cytometry was performed on a BD FACSCan™ system.
BD Pharmingen™ PE Rat Anti-Mouse CD135
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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.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
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Companion Products
The A2F10 monoclonal antibody specifically binds to Flk-2/Flt3 (Ly-72, CD135), a receptor protein tyrosine kinase closely related to c-kit, c-fms, and PDGF Receptor of the immunoglobulin superfamily. The Flt3 message is detected in hematopoietic stem cells and primitive progenitor cells in fetal liver, adult bone marrow, and fetal and adult thymus, as well as brain, placenta, and testis; but it is absent in more mature hematopoietic cells. In flow cytometric analysis, the A2F10 antibody recognizes Flt3-transfected Y3 cells (rat myeloma), but not the parent cell line in addition to recognizing early B lymphoid lineage cells in juvenile and adult bone marrow. A role for CD135 in the regulation of hematopoiesis is suggested by the observations that soluble Flk-2/Flt3 ligand can both stimulate proliferation of stem cell-enriched fetal liver, fetal thymus, and adult bone marrow populations and enhance their responses to other growth factors in vitro. In addition, injection of Flk-2/Flt3 ligand stimulates extramedullary hematopoiesis in the mouse spleen and accumulation of dendritic cells in the hematopoietic system. mAb A2F10.1 is reported to immunoprecipitate a 150-kDa surface protein from the murine myeloblast cell line M1, which naturally expresses CD135, and to inhibit the binding of Flk-2/Flt3 ligand to CD135.
Development References (10)
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Hannum C, Culpepper J, Campbell D, et al. Ligand for FLT3/FLK2 receptor tyrosine kinase regulates growth of haematopoietic stem cells and is encoded by variant RNAs. Nature. 1994; 368(2):643-648. (Biology). View Reference
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Lyman SD, James L, Vanden Bos T, et al. Molecular cloning of a ligand for the flt3/flk-2 tyrosine kinase receptor: a proliferative factor for primitive hematopoietic cells. Cell. 1993; 75(6):1157-1167. (Biology). View Reference
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Matthews W, Jordan CT, Wiegand GW, Pardoll D, Lemischka IR. A receptor tyrosine kinase specific to hematopoietic stem and progenitor cell-enriched populations. Cell. 1991; 65(7):1143-1152. (Biology). View Reference
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Ogawa M, Sugawara S, Kunisada T, et al. Flt3/Flk-2 and c-Kit are not essential for the proliferation of B lymphoid progenitor cells in the bone marrow of the adult mouse. Exp Hematol. 1998; 26(6):478-488. (Immunogen: Apoptosis). View Reference
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Ogawa M, ten Boekel E, Melchers F. Identification of CD19(-)B220(+)c-Kit(+)Flt3/Flk-2(+)cells as early B lymphoid precursors before pre-B-I cells in juvenile mouse bone marrow. Int Immunol. 2000; 12(3):313-324. (Biology). View Reference
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Orlic D, Fischer R, Nishikawa S, Nienhuis AW, Bodine D. Purification and characterization of heterogeneous pluripotent hematopoietic stem cell populations expressing high levels of c-kit receptor. Blood. 1993; 82(3):762-770. (Biology). View Reference
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Shurin MR, Pandharipande PP, Zorina TD, et al. FLT3 ligand induces the generation of functionally active dendritic cells in mice. Cell Immunol. 1997; 179(2):174-184. (Biology). View Reference
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Veiby OP, Jacobsen FW, Cui L, Lyman SD, Jacobsen SE. The flt3 ligand promotes the survival of primitive hemopoietic progenitor cells with myeloid as well as B lymphoid potential. Suppression of apoptosis and counteraction by TNF-alpha and TGF-beta. J Immunol. 1996; 157(7):2953-2960. (Biology). View Reference
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Veiby OP, Lyman SD, Jacobsen SE. Combined signaling through interleukin-7 receptors and flt3 but not c-kit potently and selectively promotes B-cell commitment and differentiation from uncommitted murine bone marrow progenitor cells.. Blood. 1996; 88(4):1256-65. (Biology). View Reference
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Wasserman R, Li YS, Hardy RR. Differential expression of the blk and ret tyrosine kinases during B lineage development is dependent on Ig rearrangement. J Immunol. 1995; 155(2):644-651. (Biology). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.