CD103 Antibody, anti-mouse, REAfinity™

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

CD103 Antibody, anti-mouse, REAfinity™Extended ValidationRecombinant

Clone:

REA789

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

FC, MICS, IF, IHC

Alternative names:

ITGAE, aM290, alpha-E1, alpha-M290, integrin alpha E

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

APC
Biotin
PE
PerCP-Vio 700
Vio Bright B515
VioBlue

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:

130-111-686, 130-111-608

Protocols:

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

Description:

Splenocytes of BALB/c mice were stained with CD103 antibodies or with the corresponding REA Control antibodies (left image) as well as with CD3ε antibodies. Flow cytometry was performed using the MACSQuant^® Analyzer. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.

MICS (MACSima Imaging Cyclic Staining) applications forCD103 Antibody, anti-mouse, REAfinity™

APC
PE

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

Acetone

Tested:

during development

Species tested:

mouse

Products used:

130-111-686, 130-111-608

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Extended validation for CD103 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 REA789
2E7	++
Cells were incubated with an excess of purified unconjugated CD103 (REA789) 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. Splenocytes from C57BL/6 mice 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. Splenocytes from C57BL/6 mice 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. Splenocytes from C57BL/6 mice 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. Splenocytes from C57BL/6 mice 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. Splenocytes from C57BL/6 mice 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 (REA789). 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 (REA789). 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-mouse, REAfinity™

Overview

Clone REA789 recognizes the mouse CD103 antigen, a 175 kDa large, type I transmembrane glycoprotein also known as αE integrin. CD103 associates non-covalently with integrin β7 thereby forming the heterodimeric integrin αEβ7. It mediates cell-cell contact and is involved in homing processes by binding to its ligand e-cadherin. CD103 is expressed on >90% of intestinal intraepithelial lymphocytes (IEL), epithelial T cells, and regulatory T cells. Due to its differential expression on dendritic cells from peripheral tissues, it is useful in identifying dendritic cell subsets from intestinal lamina propria, lung, and dermis.

Additional information: Clone REA789 displays negligible binding to Fc receptors

Alternative names

ITGAE, aM290, alpha-E1, alpha-M290, integrin alpha E

Detailed product information

Technical specifications

Clone	REA789
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	CD103
Alternative names of antigen	ITGAE, aM290, alpha-E1, alpha-M290, integrin alpha E
Molecular mass of antigen [kDa]	127
Distribution of antigen	B cells, T cells, lymphocytes, leukemia cells, thymocytes, breast, pancreas, skin, small intestine, spleen
Entrez Gene ID	16407
RRID	AB_2654366, AB_2654367, AB_2654368, AB_2654369, AB_2654370, AB_2654371, AB_2654372, AB_2654373, AB_2654374, AB_2904659, AB_2654365

Resources for CD103 Antibody, anti-mouse, REAfinity™

Certificates

Please follow this

link

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

References for CD103 Antibody, anti-mouse, REAfinity™

Publications

Sung, S. S. et al. (2006) A major lung CD103 alphaE-beta7 integrin-positive epithelial dendritic cell population expressing Langerin and tight junction proteins. J. Immunol. 176(9): 2161-2172
Bogunovic, M. et al. (2009) Origin of the lamina propria dendritic cell network. Immunity 31(3): 513-525
Ginhoux, F. et al. (2007)
Blood-derived dermal Langerin
⁺
dendritic cells survey the skin in the steady state.
J. Exp. Med. 204(13): 3133-3146

Applications

Extended validation