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Alexa Fluor® 488 Rat Anti-Mouse Siglec-F
Alexa Fluor® 488 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 either Alexa Fluor® 488 Rat IgG2a, κ Isotype Control (Cat. No. 557676; Left Plot) or Alexa Fluor® 488 Rat Anti-Mouse Siglec-F (Cat. No. 567005; Right Plot) at 1 µg/test. 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.
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 either Alexa Fluor® 488 Rat IgG2a, κ Isotype Control (Cat. No. 557676; Left Plot) or Alexa Fluor® 488 Rat Anti-Mouse Siglec-F (Cat. No. 567005; Right Plot) at 1 µg/test. 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™
Siglecf; Siglec5; sialic acid binding Ig-like lectin 5; SIGL5; 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)
0.2 mg/ml
AB_2870001
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 Alexa Fluor® 488 under optimum conditions, and unreacted Alexa Fluor® 488 was removed.

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.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  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.
  4. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
  5. Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
  6. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. 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 in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
  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.
567005 Rev. 1
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.

        

567005 Rev. 1
Format Details
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Alexa Fluor™ 488
Alexa Fluor™ 488 Dye is part of the BD blue family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 494-nm and an emission maximum (Em Max) at 517-nm. Alexa Fluor™ 488 is designed to be excited by the Blue laser (488 nm) and detected using an optical filter centered near 520-nm (e.g., a 530/30-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 488
Blue 488 nm
494 nm
517 nm
567005 Rev.1
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: Flow cytometry, Fluorescence activated cell sorting). View Reference
  2. Crocker PR, Varki A. Siglecs, sialic acids and innate immunity. Trends Immunol. 2001; 22(6):337-342. (Biology). View Reference
567005 Rev. 1

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.