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CD133 expression on retinoblastoma cell line and peripheral blood mononuclear cells (PBMC) Panel 1. Cells from the WERI-Rb-1 (Retinoblastoma, ATCC HTB-169) cell line were stained with either Alexa Fluor® 488 Mouse IgG2b, κ Isotype Control (Cat. No. 565383; dashed line histogram) or Alexa Fluor™ 488 Mouse Anti-Human CD133 antibody (Cat. No. 567907/567908 solid line histogram) at 0.5 μg/test. The fluorescence histogram showing CD133 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable cells. Panel 2A. Human peripheral blood mononuclear cells (PBMC) were stained with BD Horizon™ CD34 APC-R700 antibody (Cat. No. 659114) and either Alexa Fluor® 488 Mouse IgG2b, κ Isotype Control (Left Plot) or Alexa Fluor™ 488 Mouse Anti-Human CD133 antibody (Right Plot) at 0.5 μg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells before analysis. The pseudocolor density plot showing the correlated expression of CD133 (or Ig Isotype control staining) versus CD34 was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD negative) peripheral blood lymphocytes (PBL). Panel 2B. Human PBMC were similarly stained with BD Horizon™ BUV395 Mouse Anti-Human CD45 antibody (Cat. No. 563791/563792) and either Alexa Fluor® 488 Mouse IgG2b, κ Isotype Control (Left Plot) or Alexa Fluor™ 488 Mouse Anti-Human CD133 antibody (Right Plot) at 0.5 μg/test followed by Cell Viability 7-AAD Solution addition before analysis. The pseudocolor density plot showing the correlated expression of CD45 versus CD133 (or Ig Isotype control staining) was similarly derived for viable PBL. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
BD Pharmingen™ Alexa Fluor™ 488 Mouse Anti-Human CD133
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
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
- 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).
- 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.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
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- Alexa Fluor™ is a trademark of Life Technologies Corporation.
Companion Products
The 293C3 monoclonal antibody specifically recognizes CD133 which is also known as Prominin-like protein 1 (PROML1), Prominin-1 (PROM1), hProminin, Hematopoietic stem cell antigen, Macular dystrophy retinal 2 (MCDR2), Stargardt disease 4 autosomal dominant (STGD4), or AC133 antigen. CD133 is an ~120 kDa five-transmembrane, glycoprotein that is encoded by PROM1 (Prominin 1) which belongs to the Prominin gene family. This single-chain, pentaspan transmembrane glycoprotein is comprised of an extracellular N-terminus with two short intracellular sequences and two long extracellular loops followed by an intracellular C-terminus. CD133 is expressed on some cells found in different tissues including the bone marrow, cord and peripheral blood, placenta, liver, pancreas, kidney, lung, retina, brain and heart. It is expressed on a variety of cell types including hematopoietic stem cells and progenitor cells, neural stem cells, developing epithelial cells, precursor endothelial cells, and retinal cells. CD133 is expressed on some cancer cells found in leukemias, melanoma and retinoblastoma. It may serve as a cancer stem cell marker in a number of brain tumors, melanoma, colon cancer, hepatocellular carcinoma, pancreatic adenocarcinoma, and prostate cancer. A mutation in PROM1 has been associated with a form of human retinal degeneration. The 293C3 antibody recognizes a different epitope than the human CD133-specific W6B3C1 antibody.
Development References (5)
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Bühring HK, Marzer A, Lammers R, Wissinger B. CD133 cluster report. In: Mason D. David Mason .. et al., ed. Leucocyte typing VII : white cell differentiation antigens : proceedings of the Seventh International Workshop and Conference held in Harrogate, United Kingdom. Oxford: Oxford University Press; 2002:622-623.
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Koerner SP, André MC, Leibold JS, et al. An Fc-optimized CD133 antibody for induction of NK cell reactivity against myeloid leukemia.. Leukemia. 2017; 31(2):459-469. (Clone-specific: Blocking, Flow cytometry). View Reference
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Lammers R, Giesert C, Grünebach F, Marxer A, Vogel W, Bühring HJ. Monoclonal antibody 9C4 recognizes epithelial cellular adhesion molecule, a cell surface antigen expressed in early steps of erythropoiesis.. Exp Hematol. 2002; 30(6):537-45. (Immunogen: Flow cytometry). View Reference
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Maw MA, Corbeil D, Koch J, et al. A frameshift mutation in prominin (mouse)-like 1 causes human retinal degeneration.. Hum Mol Genet. 2000; 9(1):27-34. (Biology). View Reference
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Miraglia SJ, Buck D. CD133 (AC133). In: Mason D. David Mason .. et al., ed. Leucocyte typing VII : white cell differentiation antigens : proceedings of the Seventh International Workshop and Conference held in Harrogate, United Kingdom. Oxford: Oxford University Press; 2002:870-872.
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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
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.