TCRβ Antibody, anti-mouse, REAfinity™

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

TCRβ Antibody, anti-mouse, REAfinity™Extended ValidationRecombinant

Clone:

REA318

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

FC, MICS, IF, IHC

Alternative names:

TCRB, TCRbeta, Tib

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

APC
APC-Vio 770
Biotin
FITC
PE
PE-Vio 615
PE-Vio 770
PerCP-Vio 700
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:

130-123-842

Protocols:

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

Description:

Splenocytes from C57BL/6 mice were stained with Anti-TCRβ antibodies or with the corresponding REA Control antibodies (left images) as well as with CD3ε antibodies and analyzed by flow cytometry using the MACSQuant^® Analyzer. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence 4',6-diamidino-2-phenylindole (DAPI) fluorescence, as in the case of tandem conjugates.

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

APC
FITC
PE

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

Acetone

Tested:

during development

Species tested:

mouse

Products used:

130-123-842

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

PFA

Tested:

during development

Species tested:

mouse

Products used:

130-123-842

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Extended validation for TCRβ 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 REA318
H57-597	++
Cells were incubated with an excess of purified unconjugated TCRβ (REA318) 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 TCRβ. Splenocytes from C57BL/6 mice were stained with TCRβ antibodies and with a suitable counterstaining. As a control, TCRβ 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 propidium iodide 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 TCRβ. Splenocytes from C57BL/6 mice were stained with TCRβ antibodies and with a suitable counterstaining. As a control, TCRβ 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 propidium iodide 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 TCRβ. Splenocytes from C57BL/6 mice were stained with TCRβ antibodies and with a suitable counterstaining. As a control, TCRβ 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 propidium iodide 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 TCRβ (REA318). 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 TCRβ (REA318). 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 TCRβ Antibody, anti-mouse, REAfinity™

Overview

Clone REA318 recognizes the mouse T cell receptor beta chain (TCRβ). The T cell receptor (TCR) is responsible for recognizing antigens bound to major histocompatibility complex (MHC) molecules. It is a disulfide-linked membrane-anchored heterodimeric glycoprotein normally consisting of the highly variable alpha and beta chains expressed as part of a complex with the invariant CD3 chain molecules. T cells expressing this receptor are referred to as TCRα/β T cells, though a minority of T cells express an alternate receptor, formed by variable gamma and delta chains, referred as TCRγ/δ T cells. TCRβ is expressed on TCRα/β T cells and thymocytes, NK1.1

⁺

thymocytes and NK-T cells.

Additional information: Clone REA318 displays negligible binding to Fc receptors.

Alternative names

TCRB, TCRbeta, Tib

Detailed product information

Technical specifications

Clone	REA318
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	TCRβ
Alternative names of antigen	TCRB, TCRbeta, Tib
Distribution of antigen	T cells
Entrez Gene ID	21577
RRID	AB_2752042, AB_2801791, AB_2857569, AB_2889766, AB_2905412, AB_2889765, AB_2889764, AB_2654023, AB_2654024, AB_2905413, AB_2922209, AB_2752053

Resources for TCRβ Antibody, anti-mouse, REAfinity™

Certificates

Please follow this

link

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

References for TCRβ Antibody, anti-mouse, REAfinity™

Publications

Robins, H. S. et al. (2009) Comprehensive assessment of T-cell receptor β-chain diversity in αβ T cells. Blood 114(19): 4099-4107
Schumm, M. et al. (2013) Depletion of T-cell receptor alpha/beta and CD19 positive cells from apheresis products with the CliniMACS device. Cytotherapy 15(10): 1253-1258
Okuno, Y. et al. (2013)
CD8
⁺
CD122
⁺
regulatory T cells contain clonally expanded cells with identical CDR3 sequences of the T-cell receptor β-chain.
Immunology 139(3): 309-317

Applications

Extended validation