Clone:
REA197
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC
Alternative names:
Siglec-1, SN, Sialic acid-binding Ig-like lectin 1 (Siglec-1)

Extended validation for CD169 (Siglec-1) Antibody, anti-mouse, REAfinity™

Specificity

Epitope competition
In order to compare the epitope specificity of an antibody, the clone being used is compared with other known clones recognizing the same antigen in a competition assay.
Other clonesOverlap in epitope recognition with REA197
3D6.112++
Ser-4-
645608-
Cells were incubated with an excess of purified unconjugated CD169 (Siglec-1) (REA197) antibody followed by staining with fluorochrome-conjugated antibodies of other known clones against the same marker. Based on the fluorescence signal obtained, the clones were identified as recognizing completely overlapping (++), partially overlapping (+), or completely different epitopes (-) of the marker.

Sensitivity

Performance comparison
Selected fluorochrome conjugated antibodies from Miltenyi Biotec were compared to commercially available hybridoma clones in flow cytometry analysis.
View details
Flow cytometric comparison of different clones for CD169 (Siglec-1). Bone marrow cells from C57BL/6 mice were stained with CD169 (Siglec-1) antibodies and with a suitable counterstaining. As a control, CD169 (Siglec-1) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD169 (Siglec-1). Bone marrow cells from C57BL/6 mice were stained with CD169 (Siglec-1) antibodies and with a suitable counterstaining. As a control, CD169 (Siglec-1) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD169 (Siglec-1). Bone marrow cells from C57BL/6 mice were stained with CD169 (Siglec-1) antibodies and with a suitable counterstaining. As a control, CD169 (Siglec-1) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD169 (Siglec-1). Bone marrow cells from C57BL/6 mice were stained with CD169 (Siglec-1) antibodies and with a suitable counterstaining. As a control, CD169 (Siglec-1) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Flow cytometric comparison of different clones for CD169 (Siglec-1). Bone marrow cells from C57BL/6 mice were stained with CD169 (Siglec-1) antibodies and with a suitable counterstaining. As a control, CD169 (Siglec-1) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Fixation data
To provide an indication on how an antibody performs after fixation of cells, in-house data on staining results before and after fixation with 3.7% formaldehyde using Miltenyi Biotec antibodies are provided. Different experimental settings may lead to different results.
No fixation
Post fixation
View details
Comparison of staining pattern on non-fixed and fixed cells using CD169 (Siglec-1) (REA197). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
View details
Comparison of staining pattern on non-fixed and fixed cells using CD169 (Siglec-1) (REA197). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
Comparison of staining pattern on non-fixed and fixed cells using CD169 (Siglec-1) (REA197). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.

Specifications for CD169 (Siglec-1) Antibody, anti-mouse, REAfinity™

Overview

Clone REA197 recognizes the mouse CD169 antigen, a Single-pass type I membrane protein also known as Sialoadhesin or Sialic acid-binding Ig-like lectin 1 (Siglec-1). CD169 is a macrophage-restricted cell surface receptor that mediates sialic-acid dependent binding to lymphocytes, including granulocytes, monocytes, natural killer cells, B-cells and CD8
+
T-cells. It is a member of the sialic acid-binding IgG-like lectin (Siglec) family of proteins characterized by affinity to specifically sialylated ligands, and under normal conditions is expressed on subsets of macrophages in secondary lymphoid tissues, such as lymph node and spleen. However, CD169
+
macrophages can also be found in a variety of pathological conditions, including (autoimmune) inflammatory infiltrates and tumors. CD169 has been shown to contribute to sialylated pathogen uptake, antigen presentation and lymphocyte proliferation, and to influence both immunity and tolerance. Additionally, CD169 positive macrophages, along with mesenchymal stem cells and β-adrenergic neurons, form the hematopoietic stem cell niche in the bone marrow. CD169
+
macrophages mediate signaling between the various cells and seem to promote hematopoietic stem cell retention to the niche.
Additional information: Clone REA197 displays negligible binding to Fc receptors.

Alternative names

Siglec-1, SN, Sialic acid-binding Ig-like lectin 1 (Siglec-1)

Detailed product information

Technical specifications

CloneREA197
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody, human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Speciesmouse
AntigenCD169 (Siglec-1)
Alternative names of antigenSiglec-1, SN, Sialic acid-binding Ig-like lectin 1 (Siglec-1)
Molecular mass of antigen [kDa]181
Distribution of antigenmacrophages, spleen
Entrez Gene ID20612
RRIDAB_2819702, AB_2819759

Resources for CD169 (Siglec-1) Antibody, anti-mouse, REAfinity™

Certificates

Please follow this
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to search for Certificates of Analysis (CoA) by lot number.

References for CD169 (Siglec-1) Antibody, anti-mouse, REAfinity™

Publications

  1. Saunderson, S. C. et al. (2014) CD169 mediates the capture of exosomes in spleen and lymph node Blood 123(2): 208-216
  2. Chow, A. et al. (2011) Bone marrow CD169+ macrophages promote the retention of hematopoietic stem and progenitor cells in the mesenchymal stem cell niche J. Exp. Med. 208(2): 261-271
  3. Crocker, P. R. et al. (1994) Sialoadhesin, a macrophage sialic acid binding receptor for haemopoietic cells with 17 immunoglobulin-like domains EMBO J. 13(19): 4490-4503

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