CD103 Antibody, anti-human, REAfinity™

Name: CD103 Antibody, anti-human, REAfinity™
Availability: InStock

CD103 Antibody, anti-human, REAfinity™Extended ValidationRecombinant

Clone:

REA803

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

FC, MC

Alternative names:

ITGAE, HUMINAE, αE integrin, alpha E integrin, integrin alpha E

Flow cytometry applications forCD103 Antibody, anti-human, REAfinity™

APC
Biotin
PE
PE-Vio 615
PE-Vio 770
Vio Bright B515
Vio Bright V423

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:

130-111-986, 130-111-833

Protocols:

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

Description:

Human peripheral blood mononuclear cells (PBMCs) were either left unstimulated (left images) or stimulated with PHA for 72 hours. Cells were then stained with CD103 antibodies and CD3 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.

Mass cytometry applications forCD103 Antibody, anti-human, REAfinity™

pure

Conjugate:

pure

Products used:

130-122-303

Extended validation for CD103 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 REA803
Ber-ACT 8	++
B-Ly7	-
Cells were incubated with an excess of purified unconjugated CD103 (REA803) 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 CD103. Human peripheral blood mononuclear cells (PBMCs) after erythrocyte lysis were stained with CD103 antibodies and with a suitable counterstaining. As a control, CD103 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 CD103. Human peripheral blood mononuclear cells (PBMCs) after erythrocyte lysis were stained with CD103 antibodies and with a suitable counterstaining. As a control, CD103 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 CD103. Human peripheral blood mononuclear cells (PBMCs) after erythrocyte lysis were stained with CD103 antibodies and with a suitable counterstaining. As a control, CD103 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 CD103. Human peripheral blood mononuclear cells (PBMCs) after erythrocyte lysis were stained with CD103 antibodies and with a suitable counterstaining. As a control, CD103 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 CD103. Human peripheral blood mononuclear cells (PBMCs) after erythrocyte lysis were stained with CD103 antibodies and with a suitable counterstaining. As a control, CD103 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 CD103 (REA803). 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 CD103 (REA803). 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 CD103 Antibody, anti-human, REAfinity™

Overview

Clone REA803 recognizes the human CD103 antigen, a type I transmembrane protein which is also known as αE integrin (ITGAE). It associates with integrin β7 to form the heterodimeric integrin molecule αEβ7 and is a receptor for the ligand E-cadherin expressed on epithelial cells. CD103 is found on intraepithelial and peripheral lymphocytes, such as regulatory T cell subpopulations.

Additional information: Clone REA803 displays negligible binding to Fc receptors.

Alternative names

ITGAE, HUMINAE, αE integrin, alpha E integrin, integrin alpha E

Detailed product information

Technical specifications

Clone	REA803
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
Antigen	CD103
Alternative names of antigen	ITGAE, HUMINAE, αE integrin, alpha E integrin, integrin alpha E
Molecular mass of antigen [kDa]	128
Distribution of antigen	T cells, epithelial cells
Entrez Gene ID	3682
RRID	AB_2654383, AB_2654384, AB_2801884, AB_2921806, AB_2921803, AB_2654376, AB_2654377, AB_2654378, AB_2654379, AB_2654380, AB_2654381, AB_2654382, AB_2783900, AB_2783899, AB_2654375

Resources for CD103 Antibody, anti-human, REAfinity™

Documents and Protocols

Scientific posters

Multicolor immunophenotyping of human tumor cells using MACSQuant® Analyzer 16

Certificates

Please follow this

link

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

References for CD103 Antibody, anti-human, REAfinity™

Publications

Kruschwitz, M. et al. (1991) Ber-ACT8: New monoclonal antibody to the mucosa lymphocyte antigen. J Clin Pathol 44: 636-645
Shaw, S. K. et al. (1994) Molecular cloning of the human mucosal lymphocyte integrin alpha E subunit. Unusual structure and restricted RNA distribution. J. Biol. Chem. 269(8): 6016-6025
Allakhverdi, Z. et al. (2006) Expression of CD103 identifies human regulatory T-cell subsets. J. Allergy Clin. Immunol. 118(6): 1342-1349

Applications

Extended validation