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      Biotin Rat Anti-Mouse Siglec-F
      Biotin Rat Anti-Mouse Siglec-F
      Two-color flow cytometric analysis of Siglec-F expression on mouse bone marrow cells. BALB/c mouse bone marrow cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with BD Horizon™ BV421 Rat Anti-Mouse CD11b antibody (Cat. No. 562605) and with either Biotin Rat IgG2a, κ Isotype Control (Cat. No. 553928; Left Plot) or Biotin Rat Anti-Mouse Siglec-F (Cat. No. 567600; Right Plot) at 0.25 µg/test. The cells were then counterstained with PE Streptavidin (Cat. No. 554061). BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. A bivariate pseudocolor density plot showing the correlated expression of Siglec-F (or Ig Isotype control staining) versus CD11b was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) bone marrow cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
      Biotin Rat Anti-Mouse Siglec-F
      Two-color flow cytometric analysis of Siglec-F expression on mouse bone marrow cells. BALB/c mouse bone marrow cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with BD Horizon™ BV421 Rat Anti-Mouse CD11b antibody (Cat. No. 562605) and with either Biotin Rat IgG2a, κ Isotype Control (Cat. No. 553928; Left Plot) or Biotin Rat Anti-Mouse Siglec-F (Cat. No. 567600; Right Plot) at 0.25 µg/test. The cells were then counterstained with PE Streptavidin (Cat. No. 554061). BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. A bivariate pseudocolor density plot showing the correlated expression of Siglec-F (or Ig Isotype control staining) versus CD11b was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) bone marrow cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
      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)
      0.5 mg/ml
      Aqueous buffered solution containing ≤0.09% sodium azide.
      RUO


      Preparation And Storage

      Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with biotin under optimum conditions, and unreacted biotin was removed.
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      Recommended Assay Procedures

      The biotinylated H32-544 mAb is useful as a detection antibody for a sandwich ELISA that measures human Blys protein when paired with purified mouse anti-human Blys (Cat. No. 558289) as capture antibody and recombinant human Blys protein as standard. The biotinylated H32-544 antibody should be titrated between 0.25 - 2 µg/ml to determine its optimal concentration. To obtain linear standard curves, doubling dilutions of recombinant human Blys protein ranging from 1.56 - 100 ng/ml are recommended. For specific methodology, see Chapter 7: ELISA for specifically measuring the levels of cytokines, chemokines, and inflammatory mediators and their receptors. 2003. Techniques for Immune Function Analysis Application Handbook 1st Edition. BD Biosciences.  For this ELISA antibody pair, no cross-reactivity has been observed with the following: human TNF, TNFRI and TNFRII. This ELISA antibody pair shows a limited cross-reactivity with mouse Blys.

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      Product Notices

      1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      2. 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.
      3. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      5. An isotype control should be used at the same concentration as the antibody of interest.
      6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
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      567600 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.

      567600 Rev. 2
      Format Details
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      Biotin
      Biotin is a ubiquitous co-factor (also known as Vitamin B7) that has many properties that make it extremely useful for molecular biology. Biotin has an extremely high affinity for the Avidin family of proteins (Kd = 10-15 M), making it the perfect tool to link two molecules. Biotin labeled antibodies can be combined with any number of Avidin-conjugated probes in order to customize an assay to a particular need. This is especially useful in the case of magnetic cell separation using streptavidin/magnetic bead conjugates, or in the case of flow cytometry using streptavidin/fluorophore conjugates.
      Biotin
      567600 Rev.2
      Citations & References
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      View product citations for antibody "567600" on CiteAb

      Development References (4)

      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: Flow cytometry, Fluorescence activated cell sorting). View Reference
      2. Bosteels C, Neyt K, Vanheerswynghels M, et al. Inflammatory Type 2 cDCs Acquire Features of cDC1s and Macrophages to Orchestrate Immunity to Respiratory Virus Infection.. Immunity. 2020; 52(6):1039-1056.e9. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
      3. Crocker PR, Varki A. Siglecs, sialic acids and innate immunity. Trends Immunol. 2001; 22(6):337-342. (Biology). View Reference
      4. Engblom C, Pfirschke C, Zilionis R, et al. Osteoblasts remotely supply lung tumors with cancer-promoting SiglecFhigh neutrophils.. Science. 2017; 358(6367):eaal5081. (Clone-specific: Flow cytometry, Immunohistochemistry). View Reference
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      567600 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.