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.
- 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.
- 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.
- 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.
- CF™ is a trademark of Biotium, Inc.
- Tandem fluorochromes contain both an energy donor and an energy acceptor. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from the donor may be observed. Additionally, multi-laser cytometers may directly excite both the donor and acceptor fluorochromes. Therefore, we recommend for every tandem conjugate, a matched individual single-stain control be acquired for generating a compensation or spectral unmixing matrix.
Companion Products
The AFS98 monoclonal antibody specifically recognizes CD115 which is also known as Colony stimulating factor 1 Receptor (CSF-1R), Macrophage colony-stimulating factor 1 receptor (M-CSFR), or c-fms. This type I transmembrane glycoprotein is a receptor tyrosine kinase (RTK) that is encoded by Csf1r which belongs to the Ig superfamily. CD115 (CSF-1R) is comprised of an extracellular ligand-binding domain that is followed by a single transmembrane segment and a split intracellular tyrosine kinase domain. This receptor is expressed on monocytes, macrophages, dendritic cells, osteoclasts, and their precursors. Colony-stimulating factor 1 (CSF-1), also known as Macrophage colony-stimulating factor (M-CSF), binds to and signals through CD115 (CSF-1R) homodimers which undergo tyrosine autophosphorylation. The activated receptors transduce intracellular signals resulting in cytoskeletal reorganization and gene expression involved in the proliferation, differentiation, and survival of CSF-1-responding cells. Through CD115 (CSF-1R), CSF-1 regulates the release of proinflammatory cytokines and other mediators from macrophages and plays a role in the bone resorption activity of osteoclasts. Interleukin-34 (IL-34) is another ligand for CD115 (CSF-1R) that can induce similar, as well as, some different biological responses by target cells that express this receptor.
Development References (5)
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Fend L, Accart N, Kintz J et al. Therapeutic effects of anti-CD115 monoclonal antibody in mouse cancer models through dual inhibition of tumor-associated macrophages and osteoclasts. PLoS ONE. 2013; 8(9):e73310. (Clone-specific: Blocking). View Reference
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Jose MD, Le Meur Y, Atkins RC, Chadban SJ. Blockade of macrophage colony-stimulating factor reduces macrophage proliferation and accumulation in renal allograft rejection.. Am J Transplant. 2003; 3(3):294-300. (Clone-specific: Blocking, Immunohistochemistry, Inhibition). View Reference
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Murayama T, Yokode M, Kataoka H, et al. Intraperitoneal administration of anti-c-fms monoclonal antibody prevents initial events of atherogenesis but does not reduce the size of advanced lesions in apolipoprotein E-deficient mice.. Circulation. 1999; 99(13):1740-6. (Clone-specific: Flow cytometry, In vivo exacerbation). View Reference
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Rothwell VM, Rohrschneider LR. Murine c-fms cDNA: cloning, sequence analysis and retroviral expression. Oncogene Res. 1987; 1(4):311-324. (Biology). View Reference
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Sudo T, Nishikawa S, Ogawa M, et al. Functional hierarchy of c-kit and c-fms in intramarrow production of CFU-M.. Oncogene. 1995; 11(12):2469-76. (Immunogen: Blocking, Flow cytometry, Fluorescence activated cell sorting, Functional assay, Immunoprecipitation, Inhibition, In vivo exacerbation, Radioimmunoassay). 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.