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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.
Product Notices
- 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.
- Researchers should determine the optimal concentration of this reagent for their individual applications.
- The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- 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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
- 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.
Companion Products
The W5C5 monoclonal antibody specifically reacts with SUSD2 (sushi domain containing 2), which is expressed on mesenchymal stem cells (MSC) and on some cancers and neoplastic cell lines. It is not detected on hematopoietic cells of the peripheral blood and bone marrow. SUSD2 is a type I transmembrane protein that may interact with galectin-1 and the potential cytokine C10orf99, through which it may affect cell growth and immune responsiveness. It has a large extracellular region that contains a sushi domain (a conserved sequence that is often associated with proteins of the complement system). SUSD2 is a marker that is useful for identifying and isolating MSC from bone marrow and endometrial tissue, and those MSC may be induced to differentiate into a wide variety of cell types. Varying levels of SUSD2 expression may correlate with either suppression or enhancement of the growth of different types of neoplastic cells, presumably via a variety of direct and indirect mechanisms.
Development References (8)
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Bühring HJ, Battula VL, Treml S, Schewe B, Kanz L, Vogel W. Novel markers for the prospective isolation of human MSC. Ann N Y Acad Sci. 2007; 1106:262-271. (Clone-specific). View Reference
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Cheng Y, Wang X, Wang P, et al. SUSD2 is frequently downregulated and functions as a tumor suppressor in RCC and lung cancer.. Tumour Biol. 2016; 37(7):9919-30. (Biology). View Reference
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Hultgren EM, Patrick ME, Evans RL, Stoos CT, Egland KA. SUSD2 promotes tumor-associated macrophage recruitment by increasing levels of MCP-1 in breast cancer.. PLoS ONE. 2017; 12(5):e0177089. (Biology). View Reference
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Masuda H, Anwar SS, Bühring HJ, Rao JR, Gargett CE. A novel marker of human endometrial mesenchymal stem-like cells.. Cell Transplant. 2012; 21(10):2201-14. (Clone-specific). View Reference
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Pan W, Cheng Y, Zhang H, et al. CSBF/C10orf99, a novel potential cytokine, inhibits colon cancer cell growth through inducing G1 arrest.. Sci Rep. 2014; 4:6812. (Biology). View Reference
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Sivasubramaniyan K, Harichandan A, Schumann S, et al. Prospective isolation of mesenchymal stem cells from human bone marrow using novel antibodies directed against Sushi domain containing 2.. Stem Cells Dev. 2013; 22(13):1944-54. (Immunogen). View Reference
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Ulrich D, Tan KS, Deane J, et al. Mesenchymal stem/stromal cells in post-menopausal endometrium.. Hum Reprod. 2014; 29(9):1895-905. (Clone-specific). View Reference
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Watson AP, Evans RL, Egland KA. Multiple functions of sushi domain containing 2 (SUSD2) in breast tumorigenesis.. Mol Cancer Res. 2013; 11(1):74-85. (Biology). View Reference
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.