Description
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.
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.
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.
