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Alexa Fluor™ 488 Mouse Anti-Human CD135
Alexa Fluor™ 488 Mouse Anti-Human CD135

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.

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.

Product Details
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BD Pharmingen™
FLK2; FLT-3; Fms-related tyrosine kinase 3; STK-1
Human (QC Testing)
Mouse BALB/c IgG1, κ
Human FLT3 Transfected Cell Line
Flow cytometry (Routinely Tested)
5 µl
VI C-27
Aqueous buffered solution containing BSA and ≤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 the dye under optimum conditions and unreacted dye was removed.

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.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  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. 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).
  6. An isotype control should be used at the same concentration as the antibody of interest.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
  9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  10. Alexa Fluor™ is a trademark of Life Technologies Corporation.
567274 Rev. 1
Antibody Details
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4G8

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.

567274 Rev. 1
Format Details
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Alexa Fluor™ 488
Alexa Fluor™ 488 Dye is part of the BD blue family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 494-nm and an emission maximum (Em Max) at 517-nm. Alexa Fluor™ 488 is designed to be excited by the Blue laser (488 nm) and detected using an optical filter centered near 520-nm (e.g., a 530/30-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 488
Blue 488 nm
494 nm
517 nm
567274 Rev.1
Citations & References
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Development References (7)

  1. 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.
  2. 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
  3. 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
  4. 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.
  5. 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
  6. 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
  7. 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
View All (7) View Less
567274 Rev. 1

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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.