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Flow cytometric analysis of CD138 (Syndecan-1) expression on U266 cells. Cells from the Human U266 (Myeloma, ATCC® TIB-196™) cell line were stained with either BD Horizon™ RB545 Mouse IgG1, κ Isotype Control (Cat. No. 569284; dashed line histogram) or BD Horizon™ RB545 Mouse Anti-Human CD138 (Syndecan-1) antibody (Cat. No. 569745/569773; 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 CD138 (Syndecan-1) expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) cells. Samples were acquired on the BD FACSymphony™ A5 SE Cell Analyzer and spectrally unmixed using FlowJo™ v10.8 software.
BD Horizon™ RB545 Mouse Anti-Human CD138 (Syndecan-1)
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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.
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- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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).
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- 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.
- 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.
- 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 MI15 monoclonal antibody specifically binds to CD138 (Syndecan-1), an 85-92 kDa single chain transmembrane protein, which is strongly expressed on multiple-myeloma-derived cell lines and malignant plasma cell populations. It is also expressed on pre-B cells, immature B cells, and plasma cells, but not on mature circulating B-lymphocytes. Syndecan-1 is a member of the transmembrane heparan sulfate proteoglycans family. It is also expressed on some non-hematopoietic cells, including embryonic mesenchymal cells, vascular smooth muscle cells, endothelial and neural cells. CD138 binds to many extracellular matrix proteins through its heparan sulfate side-chains, like fibronectin, collagen types I, III, and V, tenascin, thrombospondin, and antithrombin III. It is considered an extracellular matrix receptor that may serve as a co-receptor for fibroblast growth factor and related molecules. Monoclonal antibody MI15 blocks the binding of clone B-B4 but not clone DL-101 (other anti-syndecan-1 antibodies) by flow cytometric analysis.
Development References (6)
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Barclay NA, Brown MH, Birkeland ML, et al, ed. The Leukocyte Antigen FactsBook. San Diego, CA: Academic Press; 1997.
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Costes V, Magen V, Legouffe E, et al. The Mi15 monoclonal antibody (anti-syndecan-1) is a reliable marker for quantifying plasma cells in paraffin-embedded bone marrow biopsy specimens.. Hum Pathol. 1999; 30(12):1405-11. (Immunogen: Immunohistochemistry). View Reference
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Gattei V, Godeas C, Degan M, Rossi FM, Aldinucci D, Pinto A. Characterization of anti-CD138 monoclonal antibodies as tools for investigating the molecular polymorphism of syndecan-1 in human lymphoma cells. Br J Haematol. 1999; 104(1):152-162. (Clone-specific: Flow cytometry, Immunoprecipitation). View Reference
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Horvathova M, Gaillard JP, Liautard J, et al. Identification of novel and specific antigens of human plasma cells by mAb. In: Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995:713-714.
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Wijdenes J, Clément C, Klein B, Dore J-M. CD138 (syndecan-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.; 1997:249-252.
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Zola H. Leukocyte and stromal cell molecules : the CD markers. Hoboken, N.J.: Wiley-Liss; 2007.
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.