Clone:
7C9
Type of antibody:
Primary antibodies
Isotype:
mouse IgG1κ
Applications:
MC, FC
Alternative names:
SAF2, SIGLEC8L

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 clonesOverlap 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.
View details
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.
View details
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.
View details
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.
View details
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
View details
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.
View details
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

Clone7C9
Clonalitymonoclonal
Isotypemouse IgG1κ
Isotype controlIsotype Control Antibody, mouse IgG1
Hostmouse
Type of antibodyPrimary antibodies
Specieshuman
AntigenSiglec-8
Alternative names of antigenSAF2, SIGLEC8L
Molecular mass of antigen [kDa]52
Distribution of antigenB 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 ID27181
RRIDAB_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

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

References for Siglec-8 Antibody, anti-human

Publications

  1. 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
  2. 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
  3. Nutku, E. et al. (2003) Ligation of Siglec-8: a selective mechanism for induction of human eosinophil apoptosis. Blood 101: 5014-5020

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