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      BV421 Mouse Anti-Human Siglec-8
      BV421 Mouse Anti-Human Siglec-8
      Flow cytometric analysis of Siglec-8 expression on peripheral blood granulocytes. Whole blood was stained with PE Mouse Anti-Human CD193 (Cat. No. 561746 or 558165) with no additional stain (to show the degree of eosinophil autofluorescence; Left Plot) or costained with either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; Middle Plot) or BD OptiBuild™ BV421 Mouse Anti-Human Siglec-8 (Cat. No. 747875; Right Plot) at 0.25 μg/test. The stained cells were then treated with 1× BD Pharm Lyse™ lysing solution (Cat. No. 555899) to remove erythrocytes. The pseudocolor dot plots showing Siglec-8 [or Ig isotype control] staining or autofluorescence versus CD193 were derived from gated events with the forward and side light-scatter characteristics of viable granulocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. The above is qualification data only and does not represent a specific OptiBuild™ lot.
      BV421 Mouse Anti-Human Siglec-8
      Flow cytometric analysis of Siglec-8 expression on peripheral blood granulocytes. Whole blood was stained with PE Mouse Anti-Human CD193 (Cat. No. 561746 or 558165) with no additional stain (to show the degree of eosinophil autofluorescence; Left Plot) or costained with either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; Middle Plot) or BD OptiBuild™ BV421 Mouse Anti-Human Siglec-8 (Cat. No. 747875; Right Plot) at 0.25 μg/test. The stained cells were then treated with 1× BD Pharm Lyse™ lysing solution (Cat. No. 555899) to remove erythrocytes. The pseudocolor dot plots showing Siglec-8 [or Ig isotype control] staining or autofluorescence versus CD193 were derived from gated events with the forward and side light-scatter characteristics of viable granulocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. The above is qualification data only and does not represent a specific OptiBuild™ lot.
      Product Details
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      BD OptiBuild™
      CDw329; SAF-2; SAF2; SIGLEC-8; SIGLEC8
      Human (Tested in Development)
      Mouse IgG1, κ
      Human Siglec-8 Recombinant Protein
      Flow cytometry (Qualified)
      0.2 mg/ml
      27181
      AB_2872337
      Aqueous buffered solution containing ≤0.09% sodium azide.
      RUO


      Preparation And Storage

      Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions that minimize unconjugated dye and antibody.
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      Recommended Assay Procedures

      BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation).  When fluorochrome conjugated antibodies are bound to CompBeads, they have spectral properties very similar to cells.   However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls.  It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

      For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions.  More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

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      Product Notices

      1. The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
      2. Researchers should determine the optimal concentration of this reagent for their individual applications.
      3. An isotype control should be used at the same concentration as the antibody of interest.
      4. 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.
      5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      6. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
      7. BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
      8. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
      9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      10. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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      747875 Rev. 2
      Antibody Details
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      837535

      The 837535 monoclonal antibody specifically recognizes Sialic acid-binding Ig-like lectin 8 (Siglec-8) which is also known as Sialoadhesin family member 2 (SAF-2). Siglec-8 is a single-pass, type I transmembrane glycoprotein that is encoded by SIGLEC8 which belongs to the Sialic acid-binding Ig-like lectins (Siglecs) superfamily. Siglec-8 has an Ig-like V-set domain followed by two Ig-like C-type domains in its extracellular region. Its intracellular region contains an immunoreceptor tyrosine-based inhibitory motif (ITIM) and an immunoreceptor tyrosine-based switch motif (ITSM) which might contribute to inhibitory signal transduction upon engagement of this receptor. A short form of Siglec-8 that results from alternative splicing and which lacks most of the intracellular domain has also been identified. Siglec-8 is expressed on eosinophils and mast cells, and weakly on basophils. Siglec-8 appears to serve as an adhesion molecule which mediates cell-cell interactions through its binding to 6' sulfated sialyl Lewis X carbohydrate structures. Siglec-8 seems to be involved in signal transduction since engagement of Siglec-8 can induce apoptosis in eosinophils or inhibit mast cell degranulation.

      The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max near 407 nm and Em Max near 421 nm, BD Horizon BV421 can be excited by the violet laser (405 nm) and detected with a 450/50 nm filter. BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific BlueTM conjugates. Due to nearly identical excitation and emission properties but different spillover characteristics, BD Horizon BV421, Pacific Blue, and BD Horizon V450 cannot be used simultaneously.

      747875 Rev. 2
      Format Details
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      BV421
      The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      altImg
      BV421
      Violet 405 nm
      407 nm
      423 nm
      747875 Rev.2
      Citations & References
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      View product citations for antibody "747875" on CiteAb

      Development References (2)

      1. Floyd H, Ni J, Cornish AL, et al. Siglec-8. A novel eosinophil-specific member of the immunoglobulin superfamily.. J Biol Chem. 2000; 275(2):861-6. (Biology). View Reference
      2. Kikly KK, Bochner BS, Freeman SD, et al. Identification of SAF-2, a novel siglec expressed on eosinophils, mast cells, and basophils. J Allergy Clin Immunol. 2000; 105(6 Pt 1):1093-1100. (Biology). View Reference
      747875 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.

      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.