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Two-color flow cytometric analysis of CD138 expression on mouse bone marrow cells. Mouse bone marrow cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with APC Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 553092/561880) and either BD Horizon™ BB515 Rat IgG2a, κ Isotype Control (Cat. No. 564418, Left Panel) or BD Horizon BB515 Rat Anti-Mouse CD138 antibody (Cat. No. 564511, Right Panel). Two-color flow cytometric contour plots showing the expression of CD138 (or Ig isotype control staining) versus CD45R/B220 were derived from gated events with the forward and side light-scatter characteristics of viable bone marrow cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.


BD Horizon™ BB515 Rat Anti-Mouse CD138

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Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
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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 cells and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.
For optimal results, it is recommended to perform 2 washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescence staining, prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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The 281-2 monoclonal antibody specifically binds to the core protein of CD138 (Syndecan-1), a cell-surface, integral membrane heparan sulfate- and chondroitin sulfate-containing proteoglycan that binds to interstitial extracellular matrix molecules. Syndecan-1 is predominantly expressed on epithelial cells, where its expression correlates with normal epithelial organization. It is also expressed on B lymphocytes at specific stages during their differentiation: precursor B cells in the bone marrow, and antibody-secreting cells including plasma cells (but not mature peripheral B cells). It is thus implicated in mediating B cell-matrix interactions. CD138 expression is also regulated during embryonic development, and the molecule shows a tissue- specific structural polymorphism resulting from different post-translational modifications. The 281-2 antibody may be used to detect the differently glycosylated forms, because it reacts with the core protein. Furthermore, the mAb detects the Syndecan-1 ectodomain which is cleaved from cell surfaces by a metalloproteinase.
The antibody was conjugated to BD Horizon BB515 which is part of the BD Horizon Brilliant™ Blue family of dyes. With an Ex Max near 490 nm and an Em Max near 515 nm, BD Horizon BB515 can be excited by the blue laser (488 nm) laser and detected with a 530/30 nm filter. This dye has been exclusively developed by BD Biosciences and is up to seven times brighter than FITC with less spillover into the PE channel. Due to similar excitation and emission properties, BB515, FITC, and Alexa Fluor® 488 cannot be used simultaneously. It is not recommended to use BB515 in cocktails that include Streptavidin conjugates as it may cause high background.

Development References (10)
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Bernfield M, Kokenyesi R, Kato M, et al. Biology of the syndecans: a family of transmembrane heparan sulfate proteoglycans. Annu Rev Cell Biol. 1992; 8:365-393. (Biology). View Reference
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Driver DJ, McHeyzer-Williams LJ, Cool M, Stetson DB, McHeyzer-Williams MG. Development and maintenance of a B220- memory B cell compartment. J Immunol. 2001; 167(3):1393-1405. (Clone-specific: Flow cytometry, Fluorescence microscopy, Immunofluorescence). View Reference
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Fitzgerald ML, Wang Z, Park PW, Murphy G, Bernfield M. Shedding of syndecan-1 and -4 ectodomains is regulated by multiple signaling pathways and mediated by a TIMP-3-sensitive metalloproteinase. J Cell Biol. 2000; 148(4):811-824. (Clone-specific: Dot Blot, Fluorescence microscopy, Immunofluorescence). View Reference
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Hayashi K, Hayashi M, Jalkanen M, Firestone JH, Trelstad RL, Bernfield M. Immunocytochemistry of cell surface heparan sulfate proteoglycan in mouse tissues. A light and electron microscopic study. J Histochem Cytochem. 1987; 35(10):1079-1088. (Clone-specific: Immunohistochemistry). View Reference
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Jalkanen M, Nguyen H, Rapraeger A, Kurn N, Bernfield M. Heparan sulfate proteoglycans from mouse mammary epithelial cells: localization on the cell surface with a monoclonal antibody. J Cell Biol. 1985; 101(3):976-984. (Immunogen: Dot Blot, ELISA, Fluorescence microscopy, Immunofluorescence, Radioimmunoassay, Western blot). View Reference
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Lalor PA, Nossal GJ, Sanderson RD, McHeyzer-Williams MG. Functional and molecular characterization of single, (4-hydroxy-3-nitrophenyl)acetyl (NP)-specific, IgG1+ B cells from antibody-secreting and memory B cell pathways in the C57BL/6 immune response to NP. Eur J Immunol. 1992; 22(11):3001-3011. (Biology: Western blot). View Reference
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Sanderson RD, Lalor P, Bernfield M. B lymphocytes express and lose syndecan at specific stages of differentiation. Cell Regul. 1989; 1(1):27-35. (Clone-specific: Flow cytometry, Immunoaffinity chromatography, Immunohistochemistry, Western blot). View Reference
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Sanderson RD, Sneed TB, Young LA, Sullivan GL, Lander AD. Adhesion of B lymphoid (MPC-11) cells to type I collagen is mediated by integral membrane proteoglycan, syndecan. J Immunol. 1992; 148(12):3902-3911. (Clone-specific: Immunoaffinity chromatography, Radioimmunoassay). View Reference
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Saunders S, Jalkanen M, O'Farrell S, Bernfield M. Molecular cloning of syndecan, an integral membrane proteoglycan. J Cell Biol. 1989; 108(4):1547-1556. (Clone-specific). View Reference
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Wehrli N, Legler DF, Finke D. Changing responsiveness to chemokines allows medullary plasmablasts to leave lymph nodes. Eur J Immunol. 2001; 31(2):609-616. (Biology: Immunohistochemistry). View Reference
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