BD Pharmingen™ Alexa Fluor™ 647 Rat Anti-Mouse CD301b (Mgl2)
Clone URA-1 (RUO)
Multicolor flow cytometric analysis of CD301b (Mgl2) expression on viable 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 PE Hamster Anti-Mouse CD11c antibody (Cat. No. 553802/557401) and with either Alexa Fluor™ 647 Rat IgG2a, κ Isotype Control (Cat. No. 557690; Left Plot) or Alexa Fluor™ 647 Rat Anti-Mouse CD301b (Mgl2) antibody (Cat. No. 569533/569534; Right Plot) at 1 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 CD301b (Mgl2) [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.
Multicolor flow cytometric analysis of CD301b (Mgl2) expression on viable 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 PE Hamster Anti-Mouse CD11c antibody (Cat. No. 553802/557401) and with either Alexa Fluor™ 647 Rat IgG2a, κ Isotype Control (Cat. No. 557690; Left Plot) or Alexa Fluor™ 647 Rat Anti-Mouse CD301b (Mgl2) antibody (Cat. No. 569533/569534; Right Plot) at 1 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 CD301b (Mgl2) [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.
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 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.
- 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.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- 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.
- 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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- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Alexa Fluor™ is a trademark of Life Technologies Corporation.
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
The URA-1 monoclonal antibody specifically recognizes the extracellular domain of the mouse CD301b. CD301b is also known as macrophage galactose-type C-type lectin 2 (MGL2) and is encoded by Mgl2 (Macrophage galactose N-acetyl-galactosamine specific lectin 2). Mouse D301b (MGL2) is a ~42 kDa type II transmembrane glycoproteins that is comprised of an extracellular region with a carbohydrate recognition domain (CRD) followed by a transmembrane region and a cytoplasmic tail. CD301b is expressed on conventional dendritic cells (cDCs) and immature DCs. CD301b recognizes molecules having galactose and N-actetylgalactosamine (GalNAc) residues. CD301b is 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 monoclonal antibody specifically recognizes both CD301a and CD301b and can reportedly inhibit their binding of carbohydrate ligands.
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. (Immunogen: Blocking, ELISA, Flow cytometry, Immunohistochemistry). View Reference
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Sato K, Imai Y, Higashi N, et al. Lack of antigen-specific tissue remodeling in mice deficient in the macrophage galactose-type calcium-type lectin 1/CD301a.. Blood. 2005; 106(1):207-15. (Clone-specific: Immunohistochemistry). 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. (Biology). View Reference
Please refer to Support Documents for Quality Certificates
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.
Please refer to Support Documents for Quality Certificates
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.