Siglec-6/CD327 Antibody (767329) [Alexa Fluor® 700]

U937 human histiocytic lymphoma cell line was stained with Mouse Anti-Human Siglec‑6/CD327 Alexa Fluor® 700‑conjugated Monoclonal Antibody (Catalog # FAB2859N, filled histogram) or isotype control antibody (open histogram). View our protocol for Staining Membrane-associated Proteins.

Human peripheral blood mononuclear cells (PBMCs) were stained with Mouse Anti-Human CD19 PE-conjugated Monoclonal Antibody (Catalog # FAB4867P) and either (A) Mouse Anti-Human Siglec-6/CD327 Alexa Fluor® 700-conjugated Monoclonal Antibody (Catalog # FAB2859N) or (B) Mouse Alexa Fluor 700 Isotype Control. View our protocol for Staining Membrane-associated Proteins.

Neutrophils and mononuclear phagocytes are expanded in the alveolar airspace following LPS inhalation. a Schematic overview of study design. Solid arrows denote LPS inhalation (red) or saline inhalation control (blue). Black arrows denote blood sampling. Dashed arrows denote BAL. b Flow cytometry of leukocyte preparations from SS-BAL, LPS-BAL, and HC blood. The CD45 versus SSC plot was used to define CD45+SSChi AM, CD45loSSCmid neutrophils and CD45+SSClo mononuclear cells (see also Supplementary Fig. 1). Monocyte/macrophages and DCs were negative for lineage markers CD3, CD19, CD20, and CD56 and expressed HLA-DR. Monocyte/macrophages were divided into CD14++CD16−, CD14++CD16+, and CD14-CD16++ populations analogous to blood classical, intermediate and non-classical monocytes. DCs within the CD14-CD16− gate were divided into subsets: Axl+Siglec6+ DC5s, CD11cloCD123+ pDCs, CD1c+BTLAlo-mid DC2/3s, and BTLAhi DC1s. Plots are representative of n = 9 SS BAL and n = 10 LPS BAL. c Concentrations of neutrophil, monocyte/macrophage and DC subsets in SS-BAL and LPS BAL. Bars represent mean and lines SEM. p-values from unpaired t-tests of SS versus LPS are shown: “ns” p > 0.05, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. d Monocyte/macrophage and DC frequency in HC blood, SS-BAL and LPS-BAL as a proportion of SSClo MHC class II-expressing cells (not-including CD1c- DCs). e Concentration of selected leukocyte populations in peripheral blood at 2-h intervals following inhalation of saline (blue line) or LPS (red line). Data points show mean ± SEM for 3–5 participants. p-values from one-way ANOVA are shown. p-value representation is described in c Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31040289), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: Flow
• Clone: 767329
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Alexa Fluor 700
• Concentration: Please see the vial label for concentration. If unlisted please contact technical services.

Siglec-6/CD327 Antibody (767329) [Alexa Fluor® 700] Summary

• Immunogen: Chinese hamster ovary cell line CHO-derived recombinant human Siglec‑6/CD327
  Gln27-Val331
  Accession # NP_942142
• Specificity: Detects human Siglec‑6/CD327 in direct ELISAs.
• Source: N/A
• Isotype: IgG2a
• Clonality: Monoclonal
• Host: Mouse
• Gene: SIGLEC6
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 5 uL/10^6 cells
• Application Notes: Flow Cytometry: Please use 0.25-1 ug of conjugated antibody per 10e6 cells.

Packaging, Storage & Formulations

• Storage: Protect from light. Do not freeze.
  • 12 months from date of receipt, 2 to 8 °C as supplied.
• Buffer: Supplied in a saline solution containing BSA and Sodium Azide.
• Preservative: Sodium Azide
• Concentration: Please see the vial label for concentration. If unlisted please contact technical services.

Notes

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.

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Siglec-6/CD327 Antibody (767329) [Alexa Fluor® 700]

• CD327 antigen
• CD327
• CD33 antigen-like 1
• CD33L1
• CD33L1OBBP1CD327
• CD33L2
• CD33LCDW327
• CDw327
• OB-BP1
• Obesity-binding protein 1
• sialic acid binding Ig-like lectin 6
• sialic acid-binding Ig-like lectin 6
• Siglec6
• Siglec-6

Background

Siglecs (Sialic acid binding Ig-like Lectins) are I-type (Ig-type) lectins that belong to the Ig superfamily. They are characterized by an N-terminal Ig-like V-type domain which mediates sialic acid binding, followed by varying numbers of Ig-like C2-type domains (1-4). Eleven human Siglecs (Siglec-1 through 11) have been cloned and characterized. Within these eleven, there are at least two groups, one of which is termed the CD33-related group. CD33-related Siglecs include CD33/Siglec-3 and Siglec-5 through 11 (1, 3). To date, no Siglec has been shown to recognize any cell surface ligand other than sialic acid. This suggests that interactions with glycans containing this carbohydrate are important in mediating the biological functions of Siglecs. The cDNA of human Siglec-6 (also known as OB-BP1 and CD33L), encodes a putative 442 amino acid (aa) protein that contains a 15 aa signal peptide, a 321 aa extracellular region, a 21 aa transmembrane region (TM), and an 85 aa cytoplasmic tail (5, 6). The extracellular region contains one N-terminal V-type Ig-like domain followed by two Ig-like C2-type domains. The cytoplasmic domain has one immunoreceptor tyrosine-based inhibition motif (ITIM). At least three additional isoforms exist, all of which encode an additional 11 aa’s at the N-terminus, likely due to the utilization of an alternate start site. Two of the three isoforms also show splicing. One isoform shows a 16 aa in-frame deletion in the second C2-like domain, while the other shows a deletion of the TM and cytoplasmic region, thus potentially generating a soluble form (6-9). Siglec-6 is found on B cells and in placenta, and would seem to have a restricted specificity for the sialyl Tn antigen (6, 10).

1. Crocker, P.R. and J. Zhang (2002) Biochem. Soc. Symp. 69:83.
2. Crocker, P.R. and A. Varki (2001) Immunology 103:137.
3. Crocker, P.R. (2002) Curr. Opin. Struct. Biol. 12:609.
4. Powell, L.D. and A. Varki (1995) J. Biol. Chem. 270:14243.
5. Takei, Y. et al. (1997) Cytogenet. Cell Genet. 78:295.
6. Patel, N. et al. (1999) J. Biol. Chem. 274:22729.
7. GenBank Accession # NP_942143.
8. GenBank Accession # NP_942142.
9. GenBank Accession # NP_001236.
10. Crocker, P.R. and A. Varki (2001) Trends Immunol. 22:337.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Siglec-6/CD327 Antibody (FAB2859N)(1)

Publication using FAB2859N

• L Jardine, S Wiscombe, G Reynolds, D McDonald, A Fuller, K Green, A Filby, I Forrest, MH Ruchaud-Sp, J Scott, M Collin, M Haniffa, AJ Simpson Lipopolysaccharide inhalation recruits monocytes and dendritic cell subsets to the alveolar airspace Nat Commun, 2019-04-30;10(1):1999. 2019-04-30 [PMID: 31040289] (Flow Cytometry, Human)

Bioinformatics

• Gene Symbol: SIGLEC6
• Uniprot:
  • NP_942142()