Skip to main content Skip to navigation
Hamburger Menu
Search icon
Country Selector Country Selector
China (Chinese)
Profile Icon Profile Icon
登录/注册 UserName
产品

解决方案

发现&学习

资源&工具

Support

Search icon Search icon
Close Menu
      Alert Icon
      PE-Cy7 Rat Anti-Mouse CD135 (FLT3)
      PE-Cy7 Rat Anti-Mouse CD135 (FLT3)
      Multicolor flow cytometric analysis of CD135 expression on mouse bone-marrow leukocytes. BALB/c mouse bone-marrow cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with BD Horizon™ BUV395 Rat Anti-Mouse CD45R/B220 (Cat. No. 563793) and BD Horizon™ BUV395 Rat Anti-Mouse CD11b (Cat. No. 563553) antibodies and with either PE-Cy7 Rat IgG2a, κ Isotype Control (Cat. No. 552784; Left Plot) or PE-Cy7 Rat Anti-Mouse CD135 antibody (Cat. No. 567594; Right Plot) 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 bivariate pseudocolor density plot showing the correlated expression of CD135 (or Ig Isotype control staining) versus CD45R/B220 and CD11b was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) bone marrow cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
      PE-Cy7 Rat Anti-Mouse CD135 (FLT3)
      Multicolor flow cytometric analysis of CD135 expression on mouse bone-marrow leukocytes. BALB/c mouse bone-marrow cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with BD Horizon™ BUV395 Rat Anti-Mouse CD45R/B220 (Cat. No. 563793) and BD Horizon™ BUV395 Rat Anti-Mouse CD11b (Cat. No. 563553) antibodies and with either PE-Cy7 Rat IgG2a, κ Isotype Control (Cat. No. 552784; Left Plot) or PE-Cy7 Rat Anti-Mouse CD135 antibody (Cat. No. 567594; Right Plot) 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 bivariate pseudocolor density plot showing the correlated expression of CD135 (or Ig Isotype control staining) versus CD45R/B220 and CD11b was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) bone marrow cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
      商品详情
      Down Arrow Up Arrow


      BD Pharmingen™
      Flt3; Fms-like tyrosine kinase 3; FLT-3; FLK-2; Fetal liver kinase 2; Ly72
      Mouse (QC Testing)
      Rat WI, also known as Wistar (outbred) IgG2a, κ
      Mouse Flt-3 Transfected Cell Line
      Flow cytometry (Routinely Tested)
      0.2 mg/ml
      14255
      Aqueous buffered solution containing ≤0.09% sodium azide.
      RUO


      准备和存储

      Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unconjugated antibody and free dye were removed.
      显示较多信息 blue down arrow 显示较少信息 blue up arrow

      推荐的实验流程

      BD® CompBeads can be used as surrogates to assess fluorescence spillover (Compensation).  When fluorochrome conjugated antibodies are bound to 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 cell and CompBead to ensure that BD® CompBeads are appropriate for your specific cellular application.

      显示较多信息 blue down arrow 显示较少信息 blue up arrow

      商品通知

      1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      2. 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.
      3. 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.
      4. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      6. 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).
      7. An isotype control should be used at the same concentration as the antibody of interest.
      8. 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.
      9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      10. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
      显示较多信息 blue down arrow 显示较少信息 blue up arrow
      567594 Rev. 1
      抗体详情
      Down Arrow Up Arrow
      A2F10.1

      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.

      567594 Rev. 1
      格式详情
      Down Arrow Up Arrow
      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.
      altImg
      PE-Cy7
      Yellow-Green 488 nm, 532 nm, 561 nm
      496 nm, 566 nm
      781 nm
      567594 Rev.1
      报价单和参考
      Down Arrow Up Arrow

      研发参考 (10)

      1. 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). 查看参考
      2. 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). 查看参考
      3. 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). 查看参考
      4. 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. (Clone-specific: Immunoprecipitation, Inhibition). 查看参考
      5. 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). 查看参考
      6. 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). 查看参考
      7. 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). 查看参考
      8. 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). 查看参考
      9. 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). 查看参考
      10. 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). 查看参考
      查看所有文件 (10) 查看更少内容
      567594 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.

      Website Feedback