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    Purified Rat Anti-Mouse Siglec-F
    Purified Rat Anti-Mouse Siglec-F
    Two-color analysis of Siglec-F expression on bone-marrow myeloid cells. BALB/c bone-marrow leukocytes were stained with either purified rat IgG2a, κ isotype control mAb R35-95 (Cat. No. 553927, left panel) or purified mAb E50-2440 (right panel) in the presence of Mouse Fc Block™ (purified anti-mouse CD16/CD32 mAb 2.4G2, Cat. No. 553141/553142), followed by biotinylated anti-rat IgG2a mAb RG7/1.30 (Cat. No. 553894), then Streptavidin-PE (Cat. No. 554061) and FITC-conjugated anti-mouse CD11b (Integrin αM chain) mAb M1/70 (Cat. No. 557396/553310). Flow cytometry was performed on a FACSCalibur™ (BD Biosciences, San Jose, CA).
    Purified Rat Anti-Mouse Siglec-F
    Two-color analysis of Siglec-F expression on bone-marrow myeloid cells. BALB/c bone-marrow leukocytes were stained with either purified rat IgG2a, κ isotype control mAb R35-95 (Cat. No. 553927, left panel) or purified mAb E50-2440 (right panel) in the presence of Mouse Fc Block™ (purified anti-mouse CD16/CD32 mAb 2.4G2, Cat. No. 553141/553142), followed by biotinylated anti-rat IgG2a mAb RG7/1.30 (Cat. No. 553894), then Streptavidin-PE (Cat. No. 554061) and FITC-conjugated anti-mouse CD11b (Integrin αM chain) mAb M1/70 (Cat. No. 557396/553310). Flow cytometry was performed on a FACSCalibur™ (BD Biosciences, San Jose, CA).
    Product Details
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    BD Pharmingen™
    CD170; SIGL5; Siglec-5; Siglec5; Siglecf; sialic acid-binding Ig-like lectin 5; siglec-5
    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), ELISA, Immunofluorescence, Immunohistochemistry-frozen (Reported)
    0.5 mg/ml
    AB_394340
    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. Store undiluted at 4°C.
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    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. If Mouse Fc Block™ is used, then it is important that the second-step anti-rat IgG antibody does not cross-react with the 2.4G2 mAb (rat IgG2b, κ); we recommend use of biotinylated or FITC-conjugated anti-rat IgG2a mAb RG7/1.30 (Cat. No. 553894 or 553896, respectively).

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    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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    552125 Rev. 2
    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.

    552125 Rev. 2
    Format Details
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    Purified
    Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
    Purified
    552125 Rev.2
    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
    552125 Rev. 2

    Please refer to Support Documents for Quality Certificates


    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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