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Multicolor flow cytometric analysis of Siglec-F expression on BALB/c mouse bone marrow cells. Bone marrow cells were stained simultaneously with FITC Rat Anti-Mouse CD11b antibody (Cat. No. 553310/557396/561688) and with either BD Horizon™ PE-CF594 Rat IgG2a, κ Isotype Control (Cat. No. 562302; Left Panel) or BD Horizon™ PE-CF594 Rat Anti-Mouse Siglec-F (Cat. No. 562757; Right Panel) in the presence of Mouse Fc Block™ (purified anti-mouse CD16/CD32 mAb 2.4G2, Cat. No. 553141/553142). Two-color flow cytometric dot plots show the correlated expression patterns of Siglec-F (or Ig isotype control staining) versus CD11b for gated events with the forward and side light-scatter characteristics of viable bone marrow cells. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
Multicolor flow cytometric analysis of Siglec-F expression on BALB/c mouse bone marrow cells. Bone marrow cells were stained simultaneously with FITC Rat Anti-Mouse CD11b antibody (Cat. No. 553310/557396/561688) and with either BD Horizon™ PE-CF594 Rat IgG2a, κ Isotype Control (Cat. No. 562302; Left Panel) or BD Horizon™ PE-CF594 Rat Anti-Mouse Siglec-F (Cat. No. 562757; Right Panel) in the presence of Mouse Fc Block™ (purified anti-mouse CD16/CD32 mAb 2.4G2, Cat. No. 553141/553142). Two-color flow cytometric dot plots show the correlated expression patterns of Siglec-F (or Ig isotype control staining) versus CD11b for gated events with the forward and side light-scatter characteristics of viable bone marrow cells. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
Multicolor flow cytometric analysis of Siglec-F expression on BALB/c mouse bone marrow cells. Bone marrow cells were stained simultaneously with FITC Rat Anti-Mouse CD11b antibody (Cat. No. 553310/557396/561688) and with either BD Horizon™ PE-CF594 Rat IgG2a, κ Isotype Control (Cat. No. 562302; Left Panel) or BD Horizon™ PE-CF594 Rat Anti-Mouse Siglec-F (Cat. No. 562757; Right Panel) in the presence of Mouse Fc Block™ (purified anti-mouse CD16/CD32 mAb 2.4G2, Cat. No. 553141/553142). Two-color flow cytometric dot plots show the correlated expression patterns of Siglec-F (or Ig isotype control staining) versus CD11b for gated events with the forward and side light-scatter characteristics of viable bone marrow cells. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
Multicolor flow cytometric analysis of Siglec-F expression on BALB/c mouse bone marrow cells. Bone marrow cells were stained simultaneously with FITC Rat Anti-Mouse CD11b antibody (Cat. No. 553310/557396/561688) and with either BD Horizon™ PE-CF594 Rat IgG2a, κ Isotype Control (Cat. No. 562302; Left Panel) or BD Horizon™ PE-CF594 Rat Anti-Mouse Siglec-F (Cat. No. 562757; Right Panel) in the presence of Mouse Fc Block™ (purified anti-mouse CD16/CD32 mAb 2.4G2, Cat. No. 553141/553142). Two-color flow cytometric dot plots show the correlated expression patterns of Siglec-F (or Ig isotype control staining) versus CD11b for gated events with the forward and side light-scatter characteristics of viable bone marrow cells. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
Multicolor flow cytometric analysis of Siglec-F expression on BALB/c mouse bone marrow cells. Bone marrow cells were stained simultaneously with FITC Rat Anti-Mouse CD11b antibody (Cat. No. 553310/557396/561688) and with either BD Horizon™ PE-CF594 Rat IgG2a, κ Isotype Control (Cat. No. 562302; Left Panel) or BD Horizon™ PE-CF594 Rat Anti-Mouse Siglec-F (Cat. No. 562757; Right Panel) in the presence of Mouse Fc Block™ (purified anti-mouse CD16/CD32 mAb 2.4G2, Cat. No. 553141/553142). Two-color flow cytometric dot plots show the correlated expression patterns of Siglec-F (or Ig isotype control staining) versus CD11b for gated events with the forward and side light-scatter characteristics of viable bone marrow cells. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
Multicolor flow cytometric analysis of Siglec-F expression on BALB/c mouse bone marrow cells. Bone marrow cells were stained simultaneously with FITC Rat Anti-Mouse CD11b antibody (Cat. No. 553310/557396/561688) and with either BD Horizon™ PE-CF594 Rat IgG2a, κ Isotype Control (Cat. No. 562302; Left Panel) or BD Horizon™ PE-CF594 Rat Anti-Mouse Siglec-F (Cat. No. 562757; Right Panel) in the presence of Mouse Fc Block™ (purified anti-mouse CD16/CD32 mAb 2.4G2, Cat. No. 553141/553142). Two-color flow cytometric dot plots show the correlated expression patterns of Siglec-F (or Ig isotype control staining) versus CD11b for gated events with the forward and side light-scatter characteristics of viable bone marrow cells. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
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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.
This antibody is conjugated to BD Horizon™ PE-CF594, which has been developed exclusively by BD Biosciences as a better alternative to PE-Texas Red®. PE-CF594 excites and emits at similar wavelengths to PE-Texas Red® yet exhibits improved brightness and spectral characteristics. Due to PE having maximal absorption peaks at 496 nm and 564 nm, PE-CF594 can be excited by the blue (488-nm), green (532-nm) and yellow-green (561-nm) lasers and can be detected with the same filter set as PE-Texas Red® (eg 610/20-nm filter).
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