SPECIFICITY
The A2F10 antibody reacts with Flk-2/Flt31 (Ly-72), a receptor protein tyrosine kinase closely related to c-kit, c-fms, and PDGFR of the immunoglobulin superfamily.(2) The Flk-2/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.(2) A role for Flk-2/Flt3 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.(3,4) In flow cytometric analysis, the A2F10 antibody recognizes Flk-2/Flt3-transfected Y3 cells (but not the parent cell line) and small subpopulations of CD45R/B220-positive and CD11b-positive cells in adult bone marrow.(1) It does not cross-react with differentiated hematopoietic cells known to express c-fms or c-kit.(1) It is also reported to immunoprecipitate a 150-kDa surface protein from the murine myeloblast cell line M1, which naturally expresses Flk-2/Flt3, and it inhibits the binding of Flk-2/Flt3 ligand to Flk-2/Flt3.(1)
USAGE
The antibody has been tested by immunofluorescent staining (approx. 1µg/million cells) with flow cytometric analysis to assure specificity and reactivity. Other reported applications include immunoprecipitation and blocking of ligand binding.(1) We have been unable to demonstrate any applicability in immunohistochemical staining of acetone-fixed frozen sections.(5) Since applications vary, each investigator must determine dilutions appropriate for individual use.
REFERENCES
1. Ogawa, M. Personal communication.
2. Matthews, W., C.T. Jordan, G.W. Wiegand, D. Pardoll, and I.R. Lemischka. 1991. A receptor tyrosine kinase specific to hematopoietic stem and progenitor cell-enriched populations. Cell 65: 1143 - 1152.
3. Lyman, S.D., L. James, T.V. Bos, P. de Vries, K. Brasel, B. Gliniak, L.T. Hollingsworth, K.S. Picha, H.J. McKenna, R.R. Splett, F.A. Fletcher, E. Maraskovsky, T. Farrah, D. Foxworthe, D.E. Williams, and M.P. Beckmann. 1993. Molecular cloning of a ligand for the flt3/flk-2 tyrosine kinase receptor: A proliferative factor for primitive hematopoietic cells. Cell 75: 1157 - 1167.
4 . Hannum, C., J. Culpepper, D. Campbell, T. McClanahan, S. Zurawski, J.F. Bazan, R. Kastelein, S. Hudak, J. Wagner, J. Mattson, J. Luh, G. Duda, N. Martina, D. Peterson, S. Menon, A. Shanafelt, M. Muench, G. Kelner, R. Namikawa, D. Rennick, M.-G. Roncarolo, A. Zlotnik, O. Rosnet, P. Dubreuil, D. Birnbaum, and F. Lee. 1994. Ligand for Flt3/Flk2 receptor tyrosine kinase regulates growth of haematopoietic stem cells and is encoded by variant RNAs. Nature 368: 643 - 648.
5. PharMingen. Unpublished results.
| Cat. No. | Description | Clone | Isotype | Size |
| 09891D | Purified anti-mouse Flk-2/Flt3 | A2F10.1 | Rat IgG2a | 0.5mg |
| 09895A | PE labelled anti-mouse Flk-2/Flt3 | A2F10.1 | Rat IgG2a | 0.2mg |
PharMingen also supplies a complete line of antibodies to mouse cell surface antigens, including:
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