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PE Rat Anti-Mouse Siglec-F
PE Rat Anti-Mouse Siglec-F

Two-color analysis of Siglec-F expression on bone-marrow myeloid cells. BALB/c bone-marrow leukocytes were simultaneously stained with FITC-conjugated anti-mouse CD11b (Integrin αM chain) mAb M1/70 (Cat. No.  557396/553310) and either PE-conjugated rat IgG2a, κ isotype control mAb R35-95 (Cat. No. 553930, left panel) or PE-conjugated mAb E50-2440 (right panel) in the presence of Mouse Fc Block™ (purified anti-mouse CD16/CD32 mAb 2.4G2, Cat. No. 553141/553142). Flow cytometry was performed on a FACSCalibur™ (BD Biosciences, San Jose, CA).

Two-color analysis of Siglec-F expression on bone-marrow myeloid cells. BALB/c bone-marrow leukocytes were simultaneously stained with FITC-conjugated anti-mouse CD11b (Integrin αM chain) mAb M1/70 (Cat. No.  557396/553310) and either PE-conjugated rat IgG2a, κ isotype control mAb R35-95 (Cat. No. 553930, left panel) or PE-conjugated mAb E50-2440 (right panel) in the presence of Mouse Fc Block™ (purified anti-mouse CD16/CD32 mAb 2.4G2, Cat. No. 553141/553142). Flow cytometry was performed on a FACSCalibur™ (BD Biosciences, San Jose, CA).

Product Details
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BD Pharmingen™
Mouse (QC Testing)
Rat LOU, also known as Louvain, LOU/C, LOU/M IgG2a, κ
Mouse Siglec-F and human IgG Fc recombinant fusion protein
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_394341
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with R-PE under optimum conditions, and unconjugated antibody and free PE were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

Recommended Assay Procedures

Mouse Fc Block™ (purified anti-mouse CD16/CD32 mAb 2.4G2, Cat. No. 553141/553142) may help to reduce non-specific binding to cells bearing Fcγ receptors.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. 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.
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Antibody Details
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E50-2440

The E50-2440 monoclonal antibody specifically recognizes Siglec-F. Siglecs are the sialic acid-binding immunoglobulin superfamily lectins defined in the human, each of which has a distinctive expression pattern in the hematopoietic system and at least some of which are known to mediate cell-cell interactions. Orthologous proteins of human Siglec-1 (Sialoadhesin or CD169), Siglec-2 (CD22), and Siglec-4 (myelin-associated glycoprotein) have been characterized in the mouse. Human Siglec-3 (CD33) and Siglecs-5 through -10 are encoded by a cluster of closely related genes, and each has two cytoplasmic ITIM (Immunoreceptor Tyrosine-based Inhibitory Motifs). Similarly, mouse Siglec-F is encoded by the Siglecf gene in a syntenic cluster in the mouse, and the protein has sialic acid-binding activity and an intracytoplasmic ITIM. Its expression pattern differs from those of the human Siglec-3-related proteins in that it is found on immature cells of the myelomonocytic lineage, with reduced expression on mature neutrophils and monocytes, and not on lymphoid cells. It has been proposed that mAb E50-2440 may be used for identification of immature myelomonocytic cells in the mouse.

552126 Rev. 1
Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
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Citations & References
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Development References (2)

  1. Angata T, Hingorani R, Varki NM, Varki A. Cloning and characterization of a novel mouse Siglec, mSiglec-F: differential evolution of the mouse and human (CD33) Siglec-3-related gene clusters. J Biol Chem. 2001; 276(48):45128-45136. (Immunogen: ELISA, Immunofluorescence, Immunohistochemistry). View Reference
  2. Crocker PR, Varki A. Siglecs, sialic acids and innate immunity. Trends Immunol. 2001; 22(6):337-342. (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.