Siglec-8 Antibody, anti-human

Name: Siglec-8 Antibody, anti-human
Availability: InStock

Siglec-8 Antibody, anti-humanExtended Validation

Clone:

7C9

Type of antibody:

Primary antibodies

Isotype:

mouse IgG1κ

Applications:

MC, FC

Alternative names:

SAF2, SIGLEC8L

Flow cytometry applications forSiglec-8 Antibody, anti-human

APC
Biotin
FITC
PE
PE-Vio 770

Conjugate:

APC

Recommended dilution:

1:11

Remark:

Cells should be stained prior to fixation, if formaldehyde is used as a fixative.

Tested:

QC tested

Products used:

130-098-731, 130-098-729

Protocols:

Cell surface flow cytometry staining protocol (PBS/EDTA/BSA 1:11)

Description:

Human peripheral blood cells after erythrocyte lysis were stained with Anti-Siglec-8 antibodies as well as with CD193 antibodies and analyzed by flow cytometry using the MACSQuant^® Analyzer. CD15⁺CD16^-cells were pre-gated for the analysis. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.

Mass cytometry applications forSiglec-8 Antibody, anti-human

pure

Conjugate:

pure

Products used:

130-108-015

Extended validation for Siglec-8 Antibody, anti-human

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 7C9
REA1045	++
837535	-
Cells were incubated with an excess of purified unconjugated Siglec-8 (7C9) 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 Anti-Siglec-8. Human peripheral blood mononuclear cells (PBMCs) after erythrocyte lysis were stained with Anti-Siglec-8 antibodies and with a suitable counterstaining. As a control, Anti-Siglec-8 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 propidium iodide 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 Anti-Siglec-8. Human peripheral blood mononuclear cells (PBMCs) after erythrocyte lysis were stained with Anti-Siglec-8 antibodies and with a suitable counterstaining. As a control, Anti-Siglec-8 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 propidium iodide 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 Anti-Siglec-8. Human peripheral blood mononuclear cells (PBMCs) after erythrocyte lysis were stained with Anti-Siglec-8 antibodies and with a suitable counterstaining. As a control, Anti-Siglec-8 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 propidium iodide 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 Anti-Siglec-8. Human peripheral blood mononuclear cells (PBMCs) after erythrocyte lysis were stained with Anti-Siglec-8 antibodies and with a suitable counterstaining. As a control, Anti-Siglec-8 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 propidium iodide 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-8 (7C9). 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-8 (7C9). 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-8 Antibody, anti-human

Overview

The monoclonal antibody 7C9 recognizes siglec-8, a 54 kDa type I transmembrane protein, which is a member of the Ig superfamily. Though it is mainly expressed by eosinophils, expression of siglec-8 is also found on basophils and mast cells. It has been reported that siglec-8 cross-linking with antibodies reduces eosinophil viability through induction of apoptosis.

Alternative names

SAF2, SIGLEC8L

Detailed product information

Technical specifications

Clone	7C9
Clonality	monoclonal
Isotype	mouse IgG1κ
Isotype control	Isotype Control Antibody, mouse IgG1
Host	mouse
Type of antibody	Primary antibodies
Species	human
Antigen	Siglec-8
Alternative names of antigen	SAF2, SIGLEC8L
Molecular mass of antigen [kDa]	52
Distribution of antigen	B cells, monocytes, granulocytes, leukocytes, NK cells, T cells, neutrophils, bone marrow, liver, placenta, spleen, B cells, monocytes, granulocytes, leukocytes, NK cells, neutrophils, bone marrow, liver, placenta, spleen
Entrez Gene ID	27181
RRID	AB_2653426, AB_2653427, AB_2653428, AB_2653429, AB_2653430, AB_2653431, AB_2653432, AB_2653433, AB_2653434, AB_2653435, AB_2653436

Resources for Siglec-8 Antibody, anti-human

Certificates

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References for Siglec-8 Antibody, anti-human

Publications

Nutku, E. et al. (2003) Ligation of Siglec-8: a selective mechanism for induction of human eosinophil apoptosis. Blood 101: 5014-5020
Hudson, S. A. et al. (2011) Developmental, malignancy-related, and cross-species analysis of eosinophil, mast cell, and basophil siglec-8 expression. J. Clin. Immunol. 31(6): 1045-1053
Kikly, K. K. et al. (2000) Identification of SAF-2, a novel siglec expressed on eosinophils, mast cells, and basophils. J. Allergy Clin. Immunol. 105: 1093-1100

Applications

Extended validation