Siglec-H Antibody, anti-mouse

Name: Siglec-H Antibody, anti-mouse
Availability: InStock

Siglec-H Antibody, anti-mouseExtended Validation

Clone:

551.3D3

Type of antibody:

Primary antibodies

Isotype:

rat IgG1κ

Applications:

FC

Alternative names:

SIGLECH

Flow cytometry applications forSiglec-H Antibody, anti-mouse

APC
Biotin
FITC
PE

Conjugate:

APC

Recommended dilution:

1:10

Remark:

The antibody is suited for staining of formaldehyde-fixed cells.

Tested:

QC tested

Products used:

130-102-232

Protocols:

Cell surface flow cytometry staining protocol (PBS/EDTA/BSA 1:10)

Description:

Mouse spleen cells were stained with Anti-Siglec-H antibodies conjugated to FITC (A), PE (B), APC (C) or Biotin (D), as well as with Anti-mPDCA-1-APC or Anti-mPDCA-1-PE and analyzed by flow cytometry. Cells stained with Anti-Siglec-H-Biotin were stained with Anti-Biotin-APC in addition. Cell debris and dead cells were excluded from the analysis based on scatter signals and PI fluorescence.

Extended validation for Siglec-H Antibody, anti-mouse

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 clones	Overlap in epitope recognition with 551.3D3
REA819	++
Cells were incubated with an excess of purified unconjugated Siglec-H (551.3D3) 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.

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.

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.

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

Comparison of staining pattern on non-fixed and fixed cells using Siglec-H (551.3D3). 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 (551.3D3). 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

Overview

The Anti-Siglec-H antibody reacts with the 34 kDa mouse sialic acid-binding immunoglobulin-like lectin (Siglec) H. Siglec-H is specifically expressed on mouse plasmacytoid dendritic cells

¹

- 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.

^2,3

Alternative names

SIGLECH

Detailed product information

Technical specifications

Clone	551.3D3
Clonality	monoclonal
Isotype	rat IgG1κ
Isotype control	Isotype Control Antibody, rat IgG1
Host	rat
Type of antibody	Primary antibodies
Species	mouse
Antigen	Siglec-H
Alternative names of antigen	SIGLECH
Molecular mass of antigen [kDa]	37
Entrez Gene ID	233274
RRID	AB_2660876, AB_2660877, AB_2660878, AB_2660879, AB_2660875, AB_2660874

Resources for Siglec-H Antibody, anti-mouse

Certificates

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References for Siglec-H Antibody, anti-mouse

Publications

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
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
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

Applications

Extended validation