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CD133 expression on human WERI-Rb-1 cells and human peripheral blood mononuclear cells (PBMC). Panel 1. Cells from the WERI-Rb-1 (Retinoblastoma, ATCC HTB-169) cell line were stained with either PE-CF594 Mouse IgG1, κ Isotype Control (Cat. No. 562292; dashed line histogram) or PE-CF594 Mouse Anti-Human CD133 antibody (Cat. No. 566674/566675; solid line histogram) at 0.25 µg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells before analysis. The fluorescence histogram showing CD133 expression (or Ig Isotype control staining) was derived from gated events with forward and side light-scatter characteristics of viable (7-AAD negative) cells. Panel 2A. Human PBMC were stained with BV421 Mouse Anti-Human CD34 antibody (Cat. No. 562577) and either PE-CF594 Mouse IgG1, κ Isotype Control (Left Plot) or PE-CF594 Mouse Anti-Human CD133 (Right Plot) at 0.25 µg/test. BD Via-Probe™ Cell Viability 7-AAD Solution was added to cells before analysis. Bivariate pseudocolor density plots showing the correlated expression of CD133 (or Ig Isotype control staining) versus CD34 were derived from gated events with the forward and side light-scatter characteristics of viable 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 PE-CF594 Mouse IgG1, κ Isotype Control (Left Plot) or PE-CF594 Mouse Anti-Human CD133 antibody (Right Plot) at 0.25 µg/test followed by Cell Viability 7-AAD Solution addition before analysis. Bivariate pseudocolor density plots showing the correlated expression of CD133 (or Ig Isotype control staining) versus CD45 were similarly derived for viable PBL. Flow cytometry and data analysis were performed using a BD LSRFortessa™ (Panel 1) or BD X-20 LSRFortessa™ (Panels 2A and 2B) Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
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BD Horizon™ PE-CF594 Mouse Anti-Human CD133
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BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to 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 cell and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.
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- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
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The W6B3C1 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 a variety of tissues including the bone marrow, cord and peripheral blood, placenta, liver, pancreas, kidney, lung, retina, brain and heart. It is expressed on various 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 is reportedly associated with a form of human retinal degeneration. The W6B3C1 antibody recognizes a different epitope than the human CD133-specific 293C3 antibody.
This antibody is conjugated to BD Horizon PE-CF594, which has been developed exclusively by BD Biosciences as a better alternative to PE-Texas Red®. PE-CF594 excites and emits at similar wavelengths to PE-Texas Red® yet exhibits improved brightness and spectral characteristics. Due to PE having maximal absorption peaks at 496 nm and 564 nm, PE-CF594 can be excited by the blue (488-nm), green (532-nm) and yellow-green (561-nm) lasers and can be detected with the same filter set as PE-Texas Red® (eg, 610/20-nm filter).

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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