Human Siglec-8 Alexa Fluor® 700-conjugated Antibody Summary
Met17-Ala363
Accession # Q9NYZ4
Applications
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Scientific Data
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Detection of Siglec-8 in Human Blood Granulocytes by Flow Cytometry. Human peripheral blood granulocytes were stained with Rat Anti-Human CCR3 PE-conjugated Monoclonal Antibody (Catalog # FAB155P) and either (A) Mouse Anti-Human Siglec-8 Alexa Fluor® 700-conjugated Monoclonal Antibody (Catalog # FAB7975N) or (B) Mouse IgG1Alexa Fluor 700 Isotype Control (Catalog # IC002N). View our protocol for Staining Membrane-associated Proteins.
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Preparation and Storage
- 12 months from date of receipt, 2 to 8 °C as supplied.
Background: Siglec-8
Siglec-8 (Sialic acid-binding Ig-like lectin 8), formerly called SAF-2 (sialoadhesin family member 2) and designated CD329, is a 75 kDa member of the inhibitory CD33‑related SIGLEC family within the Ig superfamily of proteins. An alternately spliced 48 kDa short form lacks most of the intracellular domain, while another isoform lacks the second of three extracellular Ig domains, representing amino acids (aa) 152-245 of the 499 aa long form. Siglec-8 is expressed exclusively on eosinophils, mast cells, and weakly on basophils, and is a marker for eosinophil terminal differentiation. It specifically recognizes 6’ sulfated sialyl Lewis X carbohydrate structures. Engagement results in eosinophils apoptosis, or mast cell inhibition of degranulation. Eosinophil Siglec F is the mouse paralog of human Siglec-8, but shares <40% amino acid sequence identity.
Product Datasheets
Product Specific Notices
This product is provided under an agreement between Life Technologies Corporation and R&D Systems, Inc, and the manufacture, use, sale or import of this product is subject to one or more US patents and corresponding non-US equivalents, owned by Life Technologies Corporation and its affiliates. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The sale of this product is expressly conditioned on the buyer not using the product or its components (1) in manufacturing; (2) to provide a service, information, or data to an unaffiliated third party for payment; (3) for therapeutic, diagnostic or prophylactic purposes; (4) to resell, sell, or otherwise transfer this product or its components to any third party, or for any other commercial purpose. Life Technologies Corporation will not assert a claim against the buyer of the infringement of the above patents based on the manufacture, use or sale of a commercial product developed in research by the buyer in which this product or its components was employed, provided that neither this product nor any of its components was used in the manufacture of such product. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, Cell Analysis Business Unit, Business Development, 29851 Willow Creek Road, Eugene, OR 97402, Tel: (541) 465-8300. Fax: (541) 335-0354.
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I used this antibody to stain human whole blood samples. First CD45 and side scatter were used to identify the major leukocyte populations in blood. Gating on granulocytes was followed by plotting with CD66b for neutrophils and Siglec-8 for eosinophils. Eosinophils can be clearly identified by higher Siglec-8 expression compared to neutrophils, which are negative for this marker. Fixation and storage at 4 degrees does not result in substantial loss of signal. This antibody was also use to create single stained compensation control using beads, but voltage may need to be optimized to achieve sufficiently bright signal as the relative brightness for AF700 is quite low.
I used the AF700 anti-Siglec 8 to identify peripheral blood eosinophils in human CF patients. This antibody has good specificity in differentiating eosinophils from other granulocytes
