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.
- 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.
- 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.
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
Companion Products
The LOM-8.7 monoclonal antibody specifically recognizes CD301a which is also known as C-type lectin domain family 10 member A (CLEC10A), Macrophage asialoglycoprotein-binding protein 1 (M-ASGP-BP-1), or Macrophage galactose N-acetyl-galactosamine specific lectin 1 (MGL1). CD301a (MGL1) is an ~42 kDa type II transmembrane glycoprotein that is encoded by Clec10a. It is comprised of an extracellular region with a carbohydrate recognition domain (CRD) followed by a transmembrane region and a cytoplasmic tail. CD301a is expressed on a subset of macrophages, conventional dendritic cells (cDCs) and plasmacytoid dendritic cells (pDCs). CD301a (MGL1) binds selectively to molecules that contain Lewis X or Lewis A structures. This receptor is involved in the recognition and endocytosis of glycoproteins and plays roles in tissue remodeling, clearance of apoptotic cells, and defense against tumor cells. The ER-MP23 monoclonal antibody recognizes both CD301a (MGL1) and its homolog, CD301b (MGL2) which is encoded by Mgl2.
Development References (3)
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Denda-Nagai K, Aida S, Saba K, et al. Distribution and function of macrophage galactose-type C-type lectin 2 (MGL2/CD301b): efficient uptake and presentation of glycosylated antigens by dendritic cells.. J Biol Chem. 2010; 285(25):19193-204. (Clone-specific: Flow cytometry, Immunohistochemistry). View Reference
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Kimura T, Imai Y, Irimura T. Calcium-dependent conformation of a mouse macrophage calcium-type lectin. Carbohydrate binding activity is stabilized by an antibody specific for a calcium-dependent epitope.. J Biol Chem. 1995; 270(27):16056-62. (Immunogen: Blocking, Flow cytometry, Immunoprecipitation). View Reference
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Tsuiji M, Fujimori M, Ohashi Y, et al. Molecular cloning and characterization of a novel mouse macrophage C-type lectin, mMGL2, which has a distinct carbohydrate specificity from mMGL1.. J Biol Chem. 2002; 277(32):28892-901. (Clone-specific: Blocking, ELISA, Immunohistochemistry). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.