IgE Antibody, anti-human, REAfinity™

Name: IgE Antibody, anti-human, REAfinity™
Availability: InStock

IgE Antibody, anti-human, REAfinity™Extended ValidationRecombinant

Clone:

REA1049

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

FC

Flow cytometry applications forIgE Antibody, anti-human, REAfinity™

APC
APC-Vio 770
Biotin
FITC
PE
PE-Vio 615
PE-Vio 770
PerCP-Vio 700
Vio Bright B515
Vio Bright R720
VioBlue
VioGreen

Conjugate:

APC

Recommended dilution:

1:50

Remark:

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

Tested:

QC tested

Products used:

130-118-003, 130-117-930

Protocols:

Cell surface flow cytometry staining protocol (PBS/EDTA/BSA 1:50)

Description:

Human peripheral blood mononuclear cells (PBMCs) were stained with Anti-IgE antibodies or with the corresponding REA Control (S) antibodies (left images) as well as with CD123 antibodies. Flow cytometry was performed using the MACSQuant^® Analyzer. The Tandem Signal Enhancer has been used to increase binding specificity of tandem-dye–conjugated antibodies. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence or 4',6-diamidino-2-phenylindole (DAPI) fluorescence, as in the case of tandem conjugates.

Extended validation for IgE 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 clones	Overlap in epitope recognition with REA1049
MHE-18	+
Ige21	-
G7-26	+
MB10-5C4	++
Cells were incubated with an excess of purified unconjugated IgE (REA1049) 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-IgE. Human peripheral blood mononuclear cells (PBMCs) were stained with Anti-IgE antibodies and with a suitable counterstaining. As a control, Anti-IgE 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 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 Anti-IgE. Human peripheral blood mononuclear cells (PBMCs) were stained with Anti-IgE antibodies and with a suitable counterstaining. As a control, Anti-IgE 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 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 Anti-IgE. Human peripheral blood mononuclear cells (PBMCs) were stained with Anti-IgE antibodies and with a suitable counterstaining. As a control, Anti-IgE 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 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 Anti-IgE. Human peripheral blood mononuclear cells (PBMCs) were stained with Anti-IgE antibodies and with a suitable counterstaining. As a control, Anti-IgE 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 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 IgE (REA1049). 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 IgE (REA1049). 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 IgE Antibody, anti-human, REAfinity™

Overview

Clone REA1049 recognizes the IgE isotype of human immunoglobulins. IgE is the shortest-lived immunoglobulin with a half-life of two days in serum. IgE plays an important role in allergy. It is especially associated with type 1 hypersensitivity, where IgE shows high-affinity binding to Fc receptors expressed on the surface of mast cells. Antigen binding to Fc receptor–bound IgE upon re-exposure to specific allergens results in degranulation and the release of a variety of mediators, such as histamine and cytokines.

IgE has two main receptors: the high-affinity receptor FcεRI, which is expressed only on mast cells and basophils, and the low-affinity receptor FcεRII (CD23), which is expressed on B cells.

Additional information: Clone REA1049 displays negligible binding to Fc receptors.

Detailed product information

Technical specifications

Clone	REA1049
Clonality	monoclonal
Isotype	recombinant human IgG1
Isotype control	REA Control Antibody (S), human IgG1
Host	human cell line
Type of antibody	Primary antibodies, Recombinant antibodies
Species	human, non-human primate
Cross-reactivity	cynomolgus monkey ( Macaca fascicularis )
Antigen	IgE
Distribution of antigen	B cells, mast cells, basophils
Entrez Gene ID	3497
RRID	AB_2733670, AB_2733862, AB_2733863, AB_2733423, AB_2733424, AB_2733960, AB_2733961, AB_2734100, AB_2734101, AB_2732996, AB_2732997, AB_2733262, AB_2733263, AB_2905272, AB_2905271, AB_2733053, AB_2733054, AB_2734037, AB_2734038, AB_2733548, AB_2733549, AB_2905274, AB_2905273, AB_2733669

Resources for IgE Antibody, anti-human, REAfinity™

Certificates

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Applications

Extended validation