Multiparameter flow cytometric analysis of CD138 expression on human peripheral blood leucocytes. Human whole blood was stained with either BD Horizon™ RY586 Mouse IgG1, κ Isotype Control (Cat. No. 568097; Left Plot) or BD Horizon RY586 Mouse Anti-Human CD138 antibody (Cat. No. 568463/568464; Right Plot). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). The bivariate pseudocolor density plot showing the correlated expression of CD138 (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals was derived from gated events with the forward and side-light scatter characteristics of intact leucocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
Flow cytometric analysis of CD138 expression on U266 cells. Cells from the Human U266 (Myeloma, ATCC® TIB-196™) cell line were stained with either BD Horizon™ RY586 Mouse IgG1, κ Isotype Control (Cat. No. 568097; dashed line histogram) or BD Horizon™ RY586 Mouse Anti-Human CD138 antibody (Cat. No. 568463/568464; solid line histogram). BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The fluorescence histogram showing CD138 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer and FlowJo™ software.
Flow cytometric analysis of CD138 expression on U266 cells. Cells from the human U266 (Myeloma, ATCC TIB-196) cell line were stained with either BD Horizon™ RY586 Mouse IgG1, κ Isotype Control (Cat. No. 568097; dashed line histogram) or BD Horizon™ RY586 Mouse Anti-Human CD138 antibody (Cat. No. 568463/568464; solid line histogram). BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The fluorescence histogram showing CD138 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer and FlowJo™ software
Multiparameter flow cytometric analysis of CD138 expression on human peripheral blood leucocytes. Human whole blood was stained with either BD Horizon™ RY586 Mouse IgG1, κ Isotype Control (Cat. No. 568097; Left Plot) or BD Horizon RY586 Mouse Anti-Human CD138 antibody (Cat. No. 568463/568464; Right Plot). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). The bivariate pseudocolor density plot showing the correlated expression of CD138 (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals was derived from gated events with the forward and side-light scatter characteristics of intact leucocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
Flow cytometric analysis of CD138 expression on U266 cells. Cells from the Human U266 (Myeloma, ATCC® TIB-196™) cell line were stained with either BD Horizon™ RY586 Mouse IgG1, κ Isotype Control (Cat. No. 568097; dashed line histogram) or BD Horizon™ RY586 Mouse Anti-Human CD138 antibody (Cat. No. 568463/568464; solid line histogram). BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The fluorescence histogram showing CD138 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer and FlowJo™ software.
Flow cytometric analysis of CD138 expression on U266 cells. Cells from the human U266 (Myeloma, ATCC TIB-196) cell line were stained with either BD Horizon™ RY586 Mouse IgG1, κ Isotype Control (Cat. No. 568097; dashed line histogram) or BD Horizon™ RY586 Mouse Anti-Human CD138 antibody (Cat. No. 568463/568464; solid line histogram). BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The fluorescence histogram showing CD138 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer and FlowJo™ software
Product Details
BD Horizon™
Syndecan; SDC; Syndecan 1; SDC1; SYND1
Human (QC Testing)
Mouse BALB/c IgG1, κ
Human U266 and XG-1 Myeloma Cell Lines
Flow cytometry (Routinely Tested)
5 µl
VI BP100, B005
6382
Aqueous buffered solution containing ≤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
- 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- CF™ is a trademark of Biotium, Inc.
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
RY586 Mouse IgG1, κ Isotype Control RUO
Size
50 µg
Cat No.
568097
Cell Viability Solution RUO
Size
500 Tests
Cat No.
555815
568463 Rev. 2
Antibody Details
MI15
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.
568463 Rev. 2
Format Details
RY586
The BD Horizon RealYellow™ 586 (RY586) Dye is part of the BD family of yellow-green dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 565-nm and an emission maximum (Em Max) at 586-nm. Driven by BD innovation, RY586 can be used on both spectral and conventional cytometers and is designed to be excited by the Yellow-Green laser (561-nm) with minimal excitation by the 488-nm Blue laser. For conventional instruments equipped with a Yellow-Green laser (561-nm), RY586 can be used as an alternative to PE and we recommend using an optical filter centered near 586-nm (eg, a 586/15-nm bandpass filter). For spectral instruments equipped with a Yellow-Green laser (561-nm), it can be used in conjunction with PE. Compared to PE, RY586 is similar in brightness, minimal spillover into Blue detectors, and increased spillover into the 610/20-nm (PE-CF594) detector. Please ensure that your instrument configuration (lasers and optical filters) is appropriate for this dye.
RY586
Yellow-Green 561 nm
564 nm
586 nm
568463 Rev.2
Citations & References
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.
568463 Rev. 2
Please refer to Support Documents for Quality Certificates
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.
