TCR Vα7.2 Antibody, anti-human, REAfinity™

Name: TCR Vα7.2 Antibody, anti-human, REAfinity™
Availability: InStock

TCR Vα7.2 Antibody, anti-human, REAfinity™Extended ValidationRecombinant

Clone:

REA179

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

FC, MICS, IF, IHC, MC

Alternative names:

TRA, IMD7, TCRA, TCRD, TRAC, TCRAV7S1, TRAV7

Flow cytometry applications forTCR Vα7.2 Antibody, anti-human, REAfinity™

APC
APC-Vio 770
Biotin
FITC
PE
PE-Vio 615
PE-Vio 770
PerCP-Vio 700
Vio Bright B515
Vio Bright R720
Vio Bright V600
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:

130-124-026, 130-123-828

Protocols:

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

Description:

Human peripheral blood mononuclear cells (PBMCs) were stained with Anti-TCR Vα7.2 antibodies as well as with CD3 antibodies and analyzed by flow cytometry using the MACSQuant^® Analyzer. The specificity of the conjugated antibodies was confirmed by blocking the binding to the ligand, using pure unconjugated antibodies (left images). 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.

MICS (MACSima Imaging Cyclic Staining) applications forTCR Vα7.2 Antibody, anti-human, REAfinity™

APC
FITC
PE

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

PFA

Tested:

during development

Species tested:

human

Products used:

130-124-026, 130-123-828

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Mass cytometry applications forTCR Vα7.2 Antibody, anti-human, REAfinity™

pure

Conjugate:

pure

Products used:

130-126-467

Extended validation for TCR Vα7.2 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 REA179
3C10	++
Cells were incubated with an excess of purified unconjugated TCR Vα7.2 (REA179) 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 TCR Vα7.2. Human peripheral blood mononuclear cells (PBMCs) were stained with TCR Vα7.2 antibodies and with a suitable counterstaining. As a control, TCR Vα7.2 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Monocytes were excluded from the analysis. 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 TCR Vα7.2. Human peripheral blood mononuclear cells (PBMCs) were stained with TCR Vα7.2 antibodies and with a suitable counterstaining. As a control, TCR Vα7.2 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Monocytes were excluded from the analysis. 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 TCR Vα7.2. Human peripheral blood mononuclear cells (PBMCs) were stained with TCR Vα7.2 antibodies and with a suitable counterstaining. As a control, TCR Vα7.2 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Monocytes were excluded from the analysis. 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 TCR Vα7.2 (REA179). 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 TCR Vα7.2 (REA179). 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 TCR Vα7.2 Antibody, anti-human, REAfinity™

Overview

Clone REA179 recognizes the TCR chain Vα7.2, which together with Jα33 segment forms the invariant T cell receptor. Human Vα7.2-Jα33 invariant T cells are preferentially localized in the gut and are often termed as mucosal-associated T (MAIT) cells. MAIT cells are restricted to MHC class I like molecules, such as MR1. Further, MAIT cells are suggested to be involved in regulation of immune response at the mucosal surfaces, which are the primary sites for pathogen entry.

Additional information: Clone REA179 displays negligible binding to Fc receptors.

Alternative names

TRA, IMD7, TCRA, TCRD, TRAC, TCRAV7S1, TRAV7

Detailed product information

Technical specifications

Clone	REA179
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	TCR Vα7.2
Alternative names of antigen	TRA, IMD7, TCRA, TCRD, TRAC, TCRAV7S1, TRAV7
Distribution of antigen	T cells
Entrez Gene ID	28686
RRID	AB_2811544, AB_2802031, AB_2802015, AB_2921763, AB_2921756, AB_2928888, AB_2928887, AB_2905418, AB_2905417, AB_2905420, AB_2905419, AB_2889770, AB_2653668, AB_2653669, AB_2653670, AB_2653671, AB_2653676, AB_2653677, AB_2928890, AB_2928889, AB_2905422, AB_2905421, AB_2922278, AB_2922272, AB_2811576

Resources for TCR Vα7.2 Antibody, anti-human, REAfinity™

Certificates

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References for TCR Vα7.2 Antibody, anti-human, REAfinity™

Publications

Treiner, E. et al. (2003) Selection of evolutionarily conserved mucosal-associated invariant T cells by MR1. Nature 422(6928): 164-169
Wang, Y. et al. (2016) Dual T cell- and B cell-intrinsic deficiency in humans with biallelic RLTPR mutations. J. Exp. Med. 213(11): 2413-2435
Qualai, J. et al. (2016) Expression of CD11c is associated with unconventional activated T cell subsets with high migratory potential. PLoS One 11(4): e0154253
Gold, M. C. and Lewinsohn, D. M. (2013) Co-dependents: MR1-restricted MAIT cells and their antimicrobial function. Nat. Rev. Microbiol. 11(1): 14-19

Applications

Extended validation