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Flow cytometric analysis of CD135 expression on human REH cells. Cells from the human REH (Acute B cell leukemia, ATCC CRL-8283) cell line were stained with either Alexa Fluor™ 488 Mouse IgG1, κ Isotype Control (Cat. No. 557720; dashed line histogram) or Alexa Fluor™ 488 Mouse Anti-Human CD135 antibody (Cat. No. 567274/567273; solid line histogram). DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The fluorescence histogram showing CD135 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) cells. Flow cytometry and data analysis was performed using a BD™ LSR II Flow Cytometer System and FlowJo™ software.
BD Pharmingen™ Alexa Fluor™ 488 Mouse Anti-Human CD135
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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.
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- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- An isotype control should be used at the same concentration as the antibody of interest.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
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The 4G8 monoclonal antibody specifically binds to CD135 which is also known as FMS-like tyrosine kinase 3 (FLT3), Fetal liver kinase 2 (FLK2) and Stem cell tyrosine kinase 1 (STK1). CD135 is a 155-160 kDa type I transmembrane glycoprotein. It belongs to subclass III of the receptor tyrosine kinase (RTK) family which includes receptors for M-CSF (CD115), SCF/c-Kit Ligand (CD117), and PDGF (CD140a and CD140b). CD135 is expressed on multipotential, myelomonocytic and primitive B-cell progenitors. The most primitive hematopoietic progenitor cells express low levels of CD135. CD135 plays the role of growth factor receptor for early hematopoietic progenitors. Reports from studies with knockout mice have shown that targeted CD135 disruption impaired the development of primitive progenitor cells of all hematopoietic lineages with significant impact on lymphopoietic precursors.
Development References (7)
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Bürhring H-J, Birnbaum D, Brasel KA, et al. CD135 (FLT3/FLK2/STK-1) Workshop Panel report. In: Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997:875-879.
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Lyman SD, James L, Johnson L. Cloning of the human homologue of the murine flt3 ligand: a growth factor for early hematopoietic progenitor cells. Blood. 1994; 83(10):2795-27801. (Biology). View Reference
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Mackarehtschian K, Hardin JD, Moore KA, Boast S, Goff SP, Lemischka IR. Targeted disruption of the flk2/flt3 gene leads to deficiencies in primitive hematopoietic progenitors. Immunity. 1995; 3(1):147-161. (Biology). View Reference
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Rappold I, Bürhring H-J. CD135 Workshop: Functional aspects and reactivity of FLT3-specific antibodies. In: Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997:879-881.
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Rappold I, Ziegler BL, Kohler I, et al. Functional and phenotypic characterization of cord blood and bone marrow subsets expressing FLT3 (CD135) receptor tyrosine kinase. Blood. 1997; 90(1):111-125. (Immunogen: ELISA, Flow cytometry, Immunoprecipitation). View Reference
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Rosnet O, Schiff C, Pébusque MJ. Human FLT3/FLK2 gene: cDNA cloning and expression in hematopoietic cells. Blood. 1993; 82(4):1110-1119. (Biology). View Reference
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Rusten LS, Lyman SD, Veiby OP, Jacobsen SE. The FLT3 ligand is a direct and potent stimulator of the growth of primitive and committed human CD34+ bone marrow progenitor cells in vitro. Blood. 1996; 87(4):1317-1325. (Biology). View Reference
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