CD23 Antibody, anti-mouse, REAfinity™

Name: CD23 Antibody, anti-mouse, REAfinity™
Availability: InStock

CD23 Antibody, anti-mouse, REAfinity™Extended ValidationRecombinant

Clone:

REA1068

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

FC, MICS, IF, IHC

Alternative names:

Fcer2a, FCE2, FCER2, Ly-42

Flow cytometry applications forCD23 Antibody, anti-mouse, REAfinity™

APC
PE
PE-Vio 770
Vio Bright B515

Conjugate:

APC

Recommended dilution:

1:50

Remark:

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

Tested:

QC tested

Products used:

130-118-761

Protocols:

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

Description:

Splenocytes from Balb/c mice were stained with CD23 antibodies or with the corresponding REA Control antibodies (left images) as well as with CD19 antibodies. Cells were analyzed by flow cytometry using the MACSQuant^® Analyzer. 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.

MICS (MACSima Imaging Cyclic Staining) applications forCD23 Antibody, anti-mouse, REAfinity™

APC
PE

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

Acetone

Tested:

during development

Species tested:

mouse

Products used:

130-118-761

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Extended validation for CD23 Antibody, anti-mouse, 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 REA1068
B3B4	++
REAL480	++
Cells were incubated with an excess of purified unconjugated CD23 (REA1068) 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 CD23. Mouse splenocytes were stained with CD23 antibodies and with a suitable counterstaining. As a control, CD23 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 CD23. Mouse splenocytes were stained with CD23 antibodies and with a suitable counterstaining. As a control, CD23 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 CD23. Mouse splenocytes were stained with CD23 antibodies and with a suitable counterstaining. As a control, CD23 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 CD23. Mouse splenocytes were stained with CD23 antibodies and with a suitable counterstaining. As a control, CD23 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 CD23. Mouse splenocytes were stained with CD23 antibodies and with a suitable counterstaining. As a control, CD23 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 CD23 (REA1068). 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 CD23 (REA1068). 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 CD23 (REA1068). 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 CD23 (REA1068). 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 CD23 Antibody, anti-mouse, REAfinity™

Overview

Clone REA1068 recognizes mouse CD23, also known as low affinity IgE Fc receptor (FcεRII), a type II integral membrane glycoprotein. CD23 is expressed on activated B cells, follicular dendritic cells of immunized mice, on a subset of splenic dendritic cells, and mature B cells, but not on CD5

⁺

B-1 cells.

Additional information: Clone REA1068 displays negligible binding to Fc receptors.

Alternative names

Fcer2a, FCE2, FCER2, Ly-42

Detailed product information

Technical specifications

Clone	REA1068
Clonality	monoclonal
Isotype	recombinant human IgG1
Isotype control	REA Control Antibody, human IgG1
Host	human cell line
Type of antibody	Primary antibodies, Recombinant antibodies
Species	mouse
Antigen	CD23
Alternative names of antigen	Fcer2a, FCE2, FCER2, Ly-42
Molecular mass of antigen [kDa]	8
Distribution of antigen	B cells, dendritic cells, mature B cells, B cell subsets
Entrez Gene ID	14127
RRID	AB_2733788, AB_2733359, AB_2733218, AB_2734014

Resources for CD23 Antibody, anti-mouse, REAfinity™

Certificates

Please follow this

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

References for CD23 Antibody, anti-mouse, REAfinity™

Publications

Rao, M. et al. (1987) Characterization of a monoclonal antibody directed against the murine B lymphocyte receptor for IgE. J Immunol 138: 1845-1851
Maeda, K. et al. (1992) Murine follicular dendritic cells and low affinity Fc receptors for IgE (Fc epsilon RII). J Immunol 148: 2340 -2347
Cho, S. W. et al. (1997)
B cell activation and Ig, especially IgE, production is inhibited by high CD23 levels
in vivo
and
in vitro.
J Immunol 180(1): 36-46

Applications

Extended validation