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
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- For U.S. patents that may apply, see bd.com/patents.
- 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.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- 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.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- An isotype control should be used at the same concentration as the antibody of interest.
- Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
Companion Products
The ME-9F1 monoclonal antibody specifically binds to mouse CD146. The CD146 adhesion molecule is a type 1 transmembrane glycoprotein and member of the immunoglobulin superfamily. CD146 is expressed by blood vessel endothelial cells and may play roles in forming intercellular junctions between endothelial cells and influencing the transendothelial migration of other cell types. CD146 is also expressed by some melanoma cell lines, NK cells and neutrophils. CD146 is not detectable on mouse monocytes, dendritic cells, T cells, NKT cells, B cells and smooth muscle cells. Increased expression of CD146 is reportedly associated with NK cell maturation and may be used to characterize different functional NK cell subsets. Activated CD146-positive mouse NK cells reportedly are less cytotoxic and secrete less IFN-γ than their CD146-negative counterparts.
Development References (4)
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Alais S, Allioli N, Pujades C, et al. HEMCAM/CD146 downregulates cell surface expression of beta1 integrins. J Cell Sci. 2001; 114(Pt 10):1847-1859. (Biology). View Reference
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Despoix N, Walzer T, Jouve N, et al. Mouse CD146/MCAM is a marker of natural killer cell maturation. Eur J Immunol. 2008; 38(10):2855-2864. (Biology). View Reference
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Schrage A, Loddenkemper C, Erben U, et al. Murine CD146 is widely expressed on endothelial cells and is recognized by the monoclonal antibody ME-9F1. Histochem Cell Biol. 2008; 129(4):441-451. (Clone-specific: Flow cytometry, Immunofluorescence, Immunohistochemistry, Immunoprecipitation). View Reference
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Yang H, Wang S, Liu Z, et al. Isolation and characterization of mouse MUC18 cDNA gene, and correlation of MUC18 expression in mouse melanoma cell lines with metastatic ability. Gene. 2001; 265(1-2):133-145. (Biology). View Reference
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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.