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RB780 Mouse Anti-Human CD138 (Syndecan-1)
RB780 Mouse Anti-Human CD138 (Syndecan-1)
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™ RB780 Mouse IgG1, κ Isotype Control (Cat. No. 568532; dashed line histogram) or BD Horizon™ RB780 Mouse Anti-Human CD138 (Syndecan-1) antibody (Cat. No. 569110/569111; 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. Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 Cell Analyzer System and FlowJo™ software.
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™ RB780 Mouse IgG1, κ Isotype Control (Cat. No. 568532; dashed line histogram) or BD Horizon™ RB780 Mouse Anti-Human CD138 (Syndecan-1) antibody (Cat. No. 569110/569111; 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. Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 Cell Analyzer System and FlowJo™ software.
Product Details
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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/test
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

  1. When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. 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).
  4. An isotype control should be used at the same concentration as the antibody of interest.
  5. 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.
  6. 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.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. 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.
  9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  10. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
569110 Rev. 2
Antibody Details
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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.

569110 Rev. 2
Format Details
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RB780
The BD Horizon RealBlue™ 780 (RB780) Dye is part of the BD family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 781-nm. Driven by BD innovation, RB780 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), RB780 can be used as an alternative to PE-Cy7 and we recommend using an optical filter centered near 780-nm (eg, a 780/60-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PE-Cy7. RB780 is on average brighter than PE-Cy7 and has minimal spillover into Yellow-Green detectors.
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RB780
Blue 488 nm
498 nm
781 nm
569110 Rev.2
Citations & References
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View product citations for antibody "569110" on CiteAb

Development References (6)

  1. Barclay NA, Brown MH, Birkeland ML, et al, ed. The Leukocyte Antigen FactsBook. San Diego, CA: Academic Press; 1997.
  2. 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
  3. 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
  4. 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.
  5. 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.
  6. Zola H. Leukocyte and stromal cell molecules : the CD markers. Hoboken, N.J.: Wiley-Liss; 2007.
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569110 Rev. 2

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.