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Flow cytometric analysis of Siglec-10 expression on human peripheral blood lymphocytes. Human whole blood was stained with Alexa Fluor® 488 Mouse Anti-Human CD19 antibody (Cat. No. 555412) and either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; Left Plot) or BD Horizon BV421 Mouse Anti-Human Siglec-10 antibody (Cat. No. 566588; Right Plot). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Two-color flow cytometric dot plots showing the correlated expression of Siglec-10 (or Ig Isotype control staining) versus CD19 were derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Flow Cytometer System.
BD Horizon™ BV421 Mouse Anti-Human Siglec-10
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Recommended Assay Procedures
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet for the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
Product Notices
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- An isotype control should be used at the same concentration as the antibody of interest.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
- BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
The 5G6 monoclonal antibody specifically recognizes Sialic acid-binding Ig-like lectin 10 (Siglec-10). Siglec-10 is a type I transmembrane glycoprotein encoded by SIGLEC10 that belongs to the SIGLEC (sialic acid binding Ig-like lectin) family within the immunoglobulin superfamily. The extracellular region of Siglec-10 contains five Ig-like domains including an N-terminal V-set domain that contains the sialic acid-binding site and four C2-set domains. Siglec-10 preferentially binds to glycoconjugates containing α2,3- or α2,6-linked sialic acids present on human cell surfaces and may function as an adhesion receptor. Its cytoplasmic region contains two immunoreceptor tyrosine-based inhibitory motifs (ITIM) that upon phosphorylation can bind the SH2 domains of several SH2-containing phosphatases. This CD33-related Siglec has a mouse ortholog designated Siglec-G. Siglec-10 is expressed on B cells, eosinophils, monocytes, and a subset of CD16+CD56- natural killer (NK) cells. It may play a role as an inhibitory receptor for leucocytes involved in innate and adaptive immunity. The 5G6 antibody reportedly did not crossreact with Siglecs 3, 5, 7, 8 and 9.
The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.
Development References (3)
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Macauley MS, Crocker PR, Paulson JC. Siglec-mediated regulation of immune cell function in disease.. Nat Rev Immunol. 2014; 14(10):653-66. (Biology). View Reference
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Munday J, Kerr S, Ni J, et al. Identification, characterization and leucocyte expression of Siglec-10, a novel human sialic acid-binding receptor.. Biochem J. 2001; 355(Pt 2):489-97. (Immunogen: ELISA, Flow cytometry). View Reference
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Pillai S, Netravali IA, Cariappa A, Mattoo H. Siglecs and immune regulation.. Annu Rev Immunol. 2012; 30:357-92. (Biology). View Reference
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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.