Clone:
REA1045
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MC
Alternative names:
SAF2, SIGLEC8L

Extended validation for Siglec-8 Antibody, anti-human, 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 REA1045
7C9++
837535-
Cells were incubated with an excess of purified unconjugated Siglec-8 (REA1045) 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 (REA1045). 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 (REA1045). 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 (REA1045). 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, REAfinity™

Overview

Clone REA1045 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.
Additional information: Clone REA1045 displays negligible binding to Fc receptors.

Alternative names

SAF2, SIGLEC8L

Detailed product information

Technical specifications

CloneREA1045
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
AntigenSiglec-8
Alternative names of antigenSAF2, SIGLEC8L
Molecular mass of antigen [kDa]52
Distribution of antigenmast cells, basophils, eosinophils
Entrez Gene ID27181
RRIDAB_2728064, AB_2728071, AB_2728065, AB_2728072, AB_2728066, AB_2728073, AB_2728067, AB_2728074, AB_2733176, AB_2733468, AB_2733469, AB_2811643, AB_2728070

Resources for Siglec-8 Antibody, anti-human, REAfinity™

Certificates

Please follow this
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References for Siglec-8 Antibody, anti-human, REAfinity™

Publications

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

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