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PE-CF594 Mouse Anti-Human CD133
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PE-CF594 Mouse Anti-Human CD133
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.
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.
Product Details
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BD Horizon™
PROM1; PROML1; prominin-1; AC133; CORD12; MCDR2; MSTP061; RP41; STGD4
Human (QC Testing)
Mouse BALB/c IgG1, κ
WERI-RB-1 Retinoblastoma Cell
Flow cytometry (Routinely Tested)
0.2 mg/ml
VII 70485
8842
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 with BD Horizon™ PE-CF594 under optimum conditions, and unconjugated antibody and free PE-CF594 were removed.

Recommended Assay Procedures

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.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. 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.
  4. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  5. When excited by the yellow-green (561-nm) laser, the fluorescence may be brighter than when excited by the blue (488-nm) laser.
  6. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using multi-laser cytometers, which may directly excite both PE and CF™594.
  7. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
  8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  9. CF™ is a trademark of Biotium, Inc.
  10. This product is provided under an Agreement between BIOTIUM and BD Biosciences. The manufacture, use, sale, offer for sale, or import of this product is subject to one or more patents or pending applications owned or licensed by Biotium, Inc. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
  11. Texas Red is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  12. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  13. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
566675 Rev. 1
Antibody Details
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W6B3C1

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).

566675 Rev. 1
Format Details
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PE-CF594
BD Horizon™ PE-CF594 dye is a part of the BD PE family of dyes. This tandem fluorochrome is comprised of a R-Phycoerythrin (PE) donor that has excitation maxima (Ex Max) of 496-nm and 566-nm and an acceptor dye with an emission maximum (Em Max) at 615-nm. PE-CF594, driven by BD innovation, is designed to be excited by the blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and detected using an optical filter centered near 615 nm (e.g., a 610/20-nm bandpass filter). The donor dye can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and the acceptor dye can be excited by the green (532-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PE-CF594
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
615 nm
566675 Rev.1
Citations & References
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View product citations for antibody "566675" on CiteAb

Development References (5)

  1. 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.
  2. 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
  3. 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
  4. 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
  5. 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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566675 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.