FoxP3 Antibody, anti-mouse, REAfinity™

Name: FoxP3 Antibody, anti-mouse, REAfinity™
Availability: InStock

FoxP3 Antibody, anti-mouse, REAfinity™Extended ValidationRecombinant

Clone:

REA788

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

ICFC, MICS, IF, IHC

Alternative names:

AIID, DIETER, IPEX, JM2, PIDX, XPID

Intracellular flow cytometry applications forFoxP3 Antibody, anti-mouse, REAfinity™

APC
PE
Vio B515
Vio R667

Conjugate:

APC

Recommended dilution:

1:50

Remark:

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

Tested:

QC tested

Products used:

130-111-679, 130-111-601

Protocols:

FoxP3 staining protocol (1:50)

Description:

Splenocytes of C57BL/6 mice were fixed and permeabilized using the FoxP3 Staining Buffer Set. Cells were then stained with Anti-FoxP3 antibodies or with the corresponding REA Control antibodies (left image) as well as with CD4 antibodies. Flow cytometry was performed using the MACSQuant^® Analyzer. Cell debris were excluded from the analysis based on scatter signals.

MICS (MACSima Imaging Cyclic Staining) applications forFoxP3 Antibody, anti-mouse, REAfinity™

APC
PE

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

PFA

Tested:

during development

Species tested:

mouse

Products used:

130-111-679, 130-111-601

Protocols:

Protocol for MICS experiments

Extended validation for FoxP3 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 REA788
150D	-
3G3	++
FJK-16s	-
MF-14	-
MF23	-
Cells were incubated with an excess of purified unconjugated FoxP3 (REA788) 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 FoxP3. Thymocytes from C57BL/6 mice were stained with a suitable counterstaining and then fixed and permeabilized using the FoxP3 Staining Buffer Set. Cells were then stained with FoxP3 antibodies. As a control, FoxP3 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris and cell doublets were excluded from the analysis based on scatter signals. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for FoxP3. Thymocytes from C57BL/6 mice were stained with a suitable counterstaining and then fixed and permeabilized using the FoxP3 Staining Buffer Set. Cells were then stained with FoxP3 antibodies. As a control, FoxP3 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris and cell doublets were excluded from the analysis based on scatter signals. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for FoxP3. Thymocytes from C57BL/6 mice were stained with a suitable counterstaining and then fixed and permeabilized using the FoxP3 Staining Buffer Set. Cells were then stained with FoxP3 antibodies. As a control, FoxP3 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris and cell doublets were excluded from the analysis based on scatter signals. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for FoxP3. Thymocytes from C57BL/6 mice were stained with a suitable counterstaining and then fixed and permeabilized using the FoxP3 Staining Buffer Set. Cells were then stained with FoxP3 antibodies. As a control, FoxP3 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris and cell doublets were excluded from the analysis based on scatter signals. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for FoxP3. Thymocytes from C57BL/6 mice were stained with a suitable counterstaining and then fixed and permeabilized using the FoxP3 Staining Buffer Set. Cells were then stained with FoxP3 antibodies. As a control, FoxP3 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris and cell doublets were excluded from the analysis based on scatter signals. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for FoxP3. Thymocytes from C57BL/6 mice were stained with a suitable counterstaining and then fixed and permeabilized using the FoxP3 Staining Buffer Set. Cells were then stained with FoxP3 antibodies. As a control, FoxP3 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris and cell doublets were excluded from the analysis based on scatter signals. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for FoxP3. Thymocytes from C57BL/6 mice were stained with a suitable counterstaining and then fixed and permeabilized using the FoxP3 Staining Buffer Set. Cells were then stained with FoxP3 antibodies. As a control, FoxP3 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris and cell doublets were excluded from the analysis based on scatter signals. The recommended titers of respective antibodies from different suppliers were used.

Specifications for FoxP3 Antibody, anti-mouse, REAfinity™

Overview

Clone REA788 recognizes the forkhead/winged-helix transcriptional regulator FoxP3, also known as forkhead box P3, scurfin, or JM2. FoxP3 is expressed pre-dominantly in regulatory T cells (Tregs) and is a major marker and functional regulator of Treg cell development and function. Mutations in the FoxP3 gene are linked to the autoimmune manifestations observed in the Scurfy mouse and humans with immune dysregulation, polyendocrinopathy, enteropathy, X-linked (IPEX) syndrome. Studies in mice have shown that FoxP3-deficient animals lack Treg cells, whereas overexpression of the FoxP3 protein leads to profound immune suppression.

Additional information: Clone REA788 displays negligible binding to Fc receptors.

Alternative names

AIID, DIETER, IPEX, JM2, PIDX, XPID

Detailed product information

Technical specifications

Clone	REA788
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	FoxP3
Alternative names of antigen	AIID, DIETER, IPEX, JM2, PIDX, XPID
Molecular mass of antigen [kDa]	47
Distribution of antigen	T cells
Entrez Gene ID	50943
RRID	AB_2651766, AB_2651767, AB_2651768, AB_2651769, AB_2651770, AB_2651771, AB_2651772, AB_2651765

Resources for FoxP3 Antibody, anti-mouse, REAfinity™

Documents and Protocols

Scientific posters

Vio® Dyes meet REAfinity™ Antibodies – antibody-fluorochrome conjugates for high-resolution multiparameter flow cytometry analysis

Certificates

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References for FoxP3 Antibody, anti-mouse, REAfinity™

Publications

Sakaguchi, S. et al. (1995) Immunologic self-tolerance maintained by activated T cells expressing IL-2 receptor α-chains (CD25). Breakdown of a single mechanism of self-tolerance causes various autoimmune diseases. J Immunol 155: 1151-1164
Hori, S. et al. (2003) Control of regulatory T cell development by the transcription factor FoxP3. Science 299: 1057-1061
Walker, M. R. et al. (2003)
Induction of FoxP3 and acquisition of T regulatory activity by stimulated human CD4
⁺
CD25
^-
T cells.
J. Clin. Invest. 112: 1437-1443
Jaeckel, E. et al. (2005) Antigen-specific FoxP3-transduced T-cells can control established type 1 diabetes. Diabetes 54: 306-310
Ziegler, S. F. (2006) FoxP3: of mice and men. Annu. Rev. Immunol. 24: 209-226
Gavin, M. A. et al. (2006) Single-cell analysis of normal and FOXP3-mutant human T cells: FOXP3 expression without regulatory T cell development. Proc. Natl. Acad. Sci. U.S.A. 103: 6659-6664
Lamprecht, B. et al. (2008) Aberrant expression of the Tʜ2 cytokine IL-21 in Hodgkin lymphoma cells regulates STAT3 signaling and attracts Treg cells via regulation of MIP-3alpha. Blood 112(8): 3339-3347

Applications

Extended validation