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Two parameter flow cytometric analysis of Siglec-5/Siglec-14 expression on human peripheral blood leucocytes. Human peripheral blood leucocytes were stained with either BD Horizon™ BB515 Mouse IgG1, κ Isotype Control (Cat No. 564416, Left Panel) or BD Horizon BB515 Mouse Anti-Human Siglec-5/Siglec-14 (Cat No. 565198; Right Panel). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). The flow cytometric contour plots show the correlated expression patterns of Siglec-5/Siglec-14 (or Ig Isotype control staining) versus side light-scatter (SSC-a) signals for intact human peripheral blood leucocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
BD Horizon™ BB515 Mouse Anti-Human Siglec-5/Siglec-14
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 results, it is recommended to perform 2 washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescent staining, prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.
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.
- 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.
- The manufacture, use, sale, offer for sale, or import of this product is subject to one or more patents or pending applications. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. Diagnostic uses require a separate license.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
The 194128 monoclonal antibody specifically binds to Siglec-5 (Sialic acid binding Ig-like lectin 5) and Siglec-14 (Sialic acid-binding Ig-like lectin 14). Siglec-5 is also known as CD170, CD33 antigen-like 2 (CD33L2) and OBBP2 (OB binding protein-2) whereas Siglec-14 is also referred to as SIG14. Both molecules are type I transmembrane glycoproteins that belong to the SIGLEC (sialic acid binding Ig-like lectin) family within the immunoglobulin superfamily. Siglec-5 can bind to alpha-2, 3- and alpha-2, 6-linked sialic acid glycoproteins and glycolipids. It is highly expressed on neutrophils and expressed at lower levels by monocytes/macrophages and dendritic cells. Siglec-5 can function as an inhibitory receptor for some cellular responses and enhance the anti-phagocytic responses by some macrophages. Siglec-14 has 99.5% amino acid sequence homology with Siglec-5 within their first two extracellular Ig domains and has a similar ligand binding preference. Siglec-14 is expressed by cells of myeloid origin and serves as an activating receptor.
The antibody was conjugated to BD Horizon BB515 which was developed exclusively by BD Biosciences. With an excitation max of 490 nm and an emission max of 515 nm, BD Horizon BB515 can be excited by the 488 nm laser and detected in a standard FITC set (eg, 530/30-nm filter). This dye provides a much brighter alternative to FITC with less spillover into the PE detector.
Development References (5)
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Angata T, Hayakawa T, Yamanaka M, Varki A, Nakamura M. Discovery of Siglec-14, a novel sialic acid receptor undergoing concerted evolution with Siglec-5 in primates. FASEB J. 2006; 20(12):1964-1973. (Biology). View Reference
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Cornish AL, Freeman S, Forbes G, et al. Characterization of siglec-5, a novel glycoprotein expressed on myeloid cells related to CD33. Blood. 1998; 92(6):2123-2132. (Biology). View Reference
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Giebel S, Nowak I, Wojnar J, et al. Association of KIR2DS4 and its variant KIR1D with leukemia. Leukemia. 2008; 22(11):2129-2130. (Biology). View Reference
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Llinas L, Lazaro A, de Salort J, Matesanz-Isabel J, Sintes J, Engel P. Expression profiles of novel cell surface molecules on B-cell subsets and plasma cells as analyzed by flow cytometry. Immunol Lett. 2011; 134(2):113-121. (Biology). View Reference
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Nguyen DH, Hurtado-Ziola N, Gagneux P, Varki A. Loss of Siglec expression on T lymphocytes during human evolution. Proc Natl Acad Sci U S A. 2006; 103(20):7765-7770. (Biology). View Reference
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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.
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