Clone:
REA819
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC
Alternative names:
SIGLECH

Extended validation for Siglec-H 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 REA819
551++
S48-791-
551.3D3+ +
Cells were incubated with an excess of purified unconjugated Siglec-H (REA819) 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 Siglec-H. C57BL/6 mouse splenoytes were stained with Siglec-Hantibodies and with a suitable counterstaining. As a control, Siglec-Hantibody 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 Siglec-H. C57BL/6 mouse splenoytes were stained with Siglec-Hantibodies and with a suitable counterstaining. As a control, Siglec-Hantibody 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 Siglec-H. C57BL/6 mouse splenoytes were stained with Siglec-Hantibodies and with a suitable counterstaining. As a control, Siglec-Hantibody 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 Siglec-H. C57BL/6 mouse splenoytes were stained with Siglec-Hantibodies and with a suitable counterstaining. As a control, Siglec-Hantibody 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 Siglec-H. C57BL/6 mouse splenoytes were stained with Siglec-Hantibodies and with a suitable counterstaining. As a control, Siglec-Hantibody 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 Siglec-H. C57BL/6 mouse splenoytes were stained with Siglec-Hantibodies and with a suitable counterstaining. As a control, Siglec-Hantibody 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 Siglec-H (REA819). 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 Siglec-H (REA819). 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 Siglec-H (REA819). 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 Siglec-H Antibody, anti-mouse, REAfinity™

Overview

Clone REA819 recognizes the mouse siglec-H antigen, a 34 kDa transmembrane protein of the Ig superfamily known as mouse sialic acid-binding immunoglobulin-like lectin. Siglec-H is specifically expressed on mouse plasmacytoid dendritic cells, a subset of CD11c
+
dendritic cells detected at low frequency in all lymphoid tissues, peripheral blood, and some non-lymphoid tissues. Binding of antibodies to siglec-H inhibits type I interferon production, which can be induced in plasmacytoid dendritic cells by DNA and RNA viruses.
Additional information: Clone REA819 displays negligible binding to Fc receptors.

Alternative names

SIGLECH

Detailed product information

Technical specifications

CloneREA819
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody, human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Speciesmouse
AntigenSiglec-H
Alternative names of antigenSIGLECH
Molecular mass of antigen [kDa]37
Distribution of antigenT cells
Entrez Gene ID233274
RRIDAB_2653457, AB_2653458, AB_2653459, AB_2653460, AB_2653461, AB_2653462, AB_2653463, AB_2653464, AB_2653465, AB_2653466, AB_2653467, AB_2653456

Resources for Siglec-H Antibody, anti-mouse, REAfinity™

Certificates

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

References for Siglec-H Antibody, anti-mouse, REAfinity™

Publications

  1. Lund, J. et al. (2003) Toll-like receptor 9–mediated recognition of herpes simplex virus-2 by plasmacytoid dendritic cells. J. Exp. Med. 198(3): 513-520
  2. Blasius, A. et al. (2004) A cell-surface molecule selectively expressed on murine natural interferon-producing cells that blocks secretion of interferon-alpha. Blood 103(11): 4201-4206
  3. Blasius, A. L. et al. (2006) Siglec-H is an IPC-specific receptor that modulates type I IFN secretion through DAP12. Blood 107(6): 2474-2476

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