Aiolos Antibody, anti-human, REAfinity™

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

Aiolos Antibody, anti-human, REAfinity™Extended ValidationRecombinant

Clone:

REA881

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

ICFC

Alternative names:

IKZF3, Znfn1a3

Intracellular flow cytometry applications forAiolos Antibody, anti-human, REAfinity™

APC
FITC
Vio B515

Conjugate:

APC

Recommended dilution:

1:50

Remark:

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

Tested:

QC tested

Products used:

130-114-418, 130-114-311

Protocols:

Intracellular flow cytometry staining protocol (Transcription Factor Staining Buffer Set (1:50)

Description:

Human peripheral blood mononuclear cells (PBMCs) were fixed and permeabilized using the FoxP3 Staining Buffer Set. Cells were then stained with Anti-Aiolos antibodies or with the corresponding REA Control (I) antibodies (left images) as well as with CD19 antibodies. Flow cytometry was performed using the MACSQuant^® Analyzer. Cell debris were excluded from the analysis based on scatter signals.

Extended validation for Aiolos 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 REA881
14C4C97	-
16D9C97	-
S50-895	++
Cells were incubated with an excess of purified unconjugated Aiolos (REA881) 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 Aiolos. Human peripheral blood mononuclear cells (PBMCs) were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the Transcription Factor Staining Buffer Set followed by intracellular staining with Aiolos antibodies. As a control, Aiolos 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 Viobility 405/520 Fixable Dye 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 Aiolos. Human peripheral blood mononuclear cells (PBMCs) were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the Transcription Factor Staining Buffer Set followed by intracellular staining with Aiolos antibodies. As a control, Aiolos 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 Viobility 405/520 Fixable Dye 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 Aiolos. Human peripheral blood mononuclear cells (PBMCs) were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the Transcription Factor Staining Buffer Set followed by intracellular staining with Aiolos antibodies. As a control, Aiolos 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 Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Specifications for Aiolos Antibody, anti-human, REAfinity™

Overview

Clone REA881 recognizes the human transcription factor aiolos, also known as IKZF3. Aiolos is a member of the ikaros family of zinc-finger proteins and forms heterodimers with other ikaros family members. It plays an essential role in the control of B lymphocyte differentiation and proliferation and in the regulation of BCL2 expression. Furthermore, it is involved in controlling apoptosis in T cells in an IL-2–dependent manner. Aiolos is found in peripheral blood leukocytes, spleen, and thymus.

Additional information: Clone REA881 displays negligible binding to Fc receptors.

Alternative names

IKZF3, Znfn1a3

Detailed product information

Technical specifications

Clone	REA881
Clonality	monoclonal
Isotype	recombinant human IgG1
Isotype control	REA Control Antibody (I), human IgG1
Host	human cell line
Type of antibody	Primary antibodies, Recombinant antibodies
Species	human
Antigen	Aiolos
Alternative names of antigen	IKZF3, Znfn1a3
Molecular mass of antigen [kDa]	58
Distribution of antigen	leukocytes, spleen
Entrez Gene ID	22806
RRID	AB_2726551, AB_2726621, AB_2726552, AB_2726622, AB_2726553, AB_2726620

Resources for Aiolos Antibody, anti-human, REAfinity™

Certificates

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References for Aiolos Antibody, anti-human, REAfinity™

Publications

Romero, F. et al. (1999) Aiolos transcription factor controls cell death in T cells by regulating Bcl-2 expression and its cellular localization. EMBO J. 18(12): 3419-3430
Caballero, R. et al. (2007) Combinatorial effects of splice variants modulate function of Aiolos. J. Cell. Sci. 120(15): 2619-2630
Billot, K. et al. (2010) Differential aiolos expression in human hematopoietic subpopulations. Leuk. Res. 34(3): 289-293

Applications

Extended validation