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APC Rat Anti-Mouse CD135
APC Rat Anti-Mouse CD135

Flow cytometric analysis of CD135 on mouse bone marrow cells.  

Bone marow cells from BALB/c mice were stained with FITC Rat Anti-Mouse CD11b (Cat. No. 553310) and FITC Rat Anti-Mouse CD45R/B220 (Cat. No. 553088) in conjunction with either a APC Rat IgG2a, κ isotype control (left panel) or with the APC Rat Anti-Mouse CD135 antibody (right panel).  Dot plots were derived from gated events based on light scattering characteristics for bone marrow cells.  Flow cytometry was performed on a BD™ LSR II flow cytometry system.

Flow cytometric analysis of CD135 on mouse bone marrow cells.  

Bone marow cells from BALB/c mice were stained with FITC Rat Anti-Mouse CD11b (Cat. No. 553310) and FITC Rat Anti-Mouse CD45R/B220 (Cat. No. 553088) in conjunction with either a APC Rat IgG2a, κ isotype control (left panel) or with the APC Rat Anti-Mouse CD135 antibody (right panel).  Dot plots were derived from gated events based on light scattering characteristics for bone marrow cells.  Flow cytometry was performed on a BD™ LSR II flow cytometry system.

Product Details
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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
AB_1727425
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

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 APC under optimum conditions, and unconjugated antibody and free APC were removed.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNe, or red diode laser.
  4. 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.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
560718 Rev. 1
Antibody Details
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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.

560718 Rev. 1
Format Details
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APC
Allophycocyanin (APC), is part of the BD family of phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 651 nm and an emission maximum (Em Max) at 660 nm. APC is designed to be excited by the Red (627-640 nm) laser and detected using an optical filter centered near 660 nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
APC
Red 627-640 nm
651 nm
660 nm
560718 Rev.1
Citations & References
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Development References (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). View Reference
  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). View Reference
  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). View Reference
  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. (Immunogen). View Reference
  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). View Reference
  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). View Reference
  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). View Reference
  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). View Reference
  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). View Reference
  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). View Reference
View All (10) View Less
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