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PE Rat Anti-Mouse CD301a/b (MGL1/2)
PE Rat Anti-Mouse CD301a/b (MGL1/2)

Multicolor flow cytometric analysis of CD301a/b (MGL1/2) expression on mouse spleen or bone marrow-derived dendritic cells. Mouse splenocytes (Left Plots) and bone marrow-derived dendritic cells (DCs) (Right Plots) 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 either PE Rat IgG2a, κ Isotype Control (Cat. No. 553930; Top Plots) or PE Rat Anti-Mouse CD301a/b (MGL1/2) antibody (Cat. No. 567050/567051; Bottom Plots) at 1 μg/test. Spleen cells were also stained with FITC Hamster Anti-Mouse CD3 (Cat. No. 553062) and FITC Rat Anti-Mouse CD19 (Cat. No. 553785) antibodies. Bivariate pseudocolor density plots showing the correlated expression of CD301a/b (MGL1/2) [or Ig Isotype control staining] versus CD11c were derived from gated events with the forward and side light-scatter characteristics of viable (CD3- and CD19-) splenocytes or 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.

Multicolor flow cytometric analysis of CD301a/b (MGL1/2) expression on mouse spleen or bone marrow-derived dendritic cells. Mouse splenocytes (Left Plots) and bone marrow-derived dendritic cells (DCs) (Right Plots) 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 either PE Rat IgG2a, κ Isotype Control (Cat. No. 553930; Top Plots) or PE Rat Anti-Mouse CD301a/b (MGL1/2) antibody (Cat. No. 567050/567051; Bottom Plots) at 1 μg/test. Spleen cells were also stained with FITC Hamster Anti-Mouse CD3 (Cat. No. 553062) and FITC Rat Anti-Mouse CD19 (Cat. No. 553785) antibodies. Bivariate pseudocolor density plots showing the correlated expression of CD301a/b (MGL1/2) [or Ig Isotype control staining] versus CD11c were derived from gated events with the forward and side light-scatter characteristics of viable (CD3- and CD19-) splenocytes or 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.

Product Details
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BD Pharmingen™
CD301a/CD301b; MGL1/MGL2; mMGL1/mMGL2
Mouse (QC Testing)
Rat LEW, also known as Lewis IgG2a, κ
Mouse Macrophage Precursor Cell Hybrids
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_2870018
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.

Recommended Assay Procedures

BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation).  When fluorochrome conjugated antibodies are bound to 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 CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. 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.
  4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
567050 Rev. 1
Antibody Details
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ER-MP23

The ER-MP23 monoclonal antibody specifically recognizes the extracellular domains of the mouse CD301a and CD301b homologs. CD301a is also known as macrophage galactose-type C-type lectin 1 (MGL1) which is encoded by Clec10a (C-type lectin domain family 10 member A). CD301b is likewise known as macrophage galactose-type C-type lectin 2 (MGL2) and is encoded by Mgl2 (Macrophage galactose N-acetyl-galactosamine specific lectin 2). Mouse CD301a and CD301b are ~42 kDa type II transmembrane glycoproteins that are each 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) whereas CD301b is expressed on cDCs and immature DCs. CD301a binds selectively to molecules that contain Lewis X or Lewis A structures. CD301b recognizes molecules having galactose and N-actetylgalactosamine (GalNAc) residues. Both receptors are involved in the recognition and endocytosis of glycoproteins and play roles in tissue remodeling, clearance of apoptotic cells, and defense against tumor cells. The ER-MP23 antibody can reportedly inhibit the binding of carbohydrate ligands to CD301a and CD301b.

        

567050 Rev. 1
Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
567050 Rev.1
Citations & References
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Development References (6)

  1. 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. (Biology). View Reference
  2. Dupasquier M, Stoitzner P, Wan H, et al. The dermal microenvironment induces the expression of the alternative activation marker CD301/mMGL in mononuclear phagocytes, independent of IL-4/IL-13 signaling.. J Leukoc Biol. 2006; 80(4):838-49. (Clone-specific: Blocking, ELISA, Flow cytometry, Immunofluorescence). View Reference
  3. Dupasquier M, Stoitzner P, van Oudenaren A, Romani N, Leenen PJ. Macrophages and dendritic cells constitute a major subpopulation of cells in the mouse dermis.. J Invest Dermatol. 2004; 123(5):876-9. (Clone-specific: Immunofluorescence). View Reference
  4. Geutskens SB, Otonkoski T, Pulkkinen MA, Drexhage HA, Leenen PJ. Macrophages in the murine pancreas and their involvement in fetal endocrine development in vitro.. J Leukoc Biol. 2005; 78(4):845-52. (Clone-specific: Immunofluorescence, Immunohistochemistry). View Reference
  5. Jansen A, Homo-Delarche F, Hooijkaas H, Leenen PJ, Dardenne M, Drexhage HA. Immunohistochemical characterization of monocytes-macrophages and dendritic cells involved in the initiation of the insulitis and beta-cell destruction in NOD mice.. Diabetes. 1994; 43(5):667-75. (Clone-specific: Immunohistochemistry). View Reference
  6. Leenen PJ, de Bruijn MF, Voerman JS, Campbell PA, van Ewijk W. Markers of mouse macrophage development detected by monoclonal antibodies. J Immunol Methods. 1994; 174(1-2):5-19. (Clone-specific). View Reference
View All (6) View Less
567050 Rev. 1

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.