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Multicolor flow cytometric analysis of CD301a (MGL1) expression on Mouse bone marrow-derived dendritic cells. Bone marrow-derived dendritic cells (DCs) from C57BL/6 mice 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 Hamster Anti-Mouse CD11c antibody (Cat. No. 550261) and with either PE Rat IgG2a, κ Isotype Control (Cat. No. 553930; Left Plot) or PE Rat Anti-Mouse CD301a (MGL1) antibody (Cat. No. 568900/568901; Right Plot) at 0.25 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of CD301a (MGL1) [or Ig Isotype control staining] versus CD11c was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) bone marrow-derived DCs. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
BD Pharmingen™ PE Rat Anti-Mouse CD301a (MGL1)
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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 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 cell 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.
- 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.
- An isotype control should be used at the same concentration as the antibody of interest.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
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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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