CD3 Antibody, anti-non-human primate, REAfinity™

Name: CD3 Antibody, anti-non-human primate, REAfinity™
Availability: InStock

CD3 Antibody, anti-non-human primate, REAfinity™Extended ValidationRecombinant

Clone:

REA994

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

FC

Alternative names:

T3

Flow cytometry applications forCD3 Antibody, anti-non-human primate, REAfinity™

APC
APC-Vio 770
Biotin
FITC
PE
PE-Vio 615
PE-Vio 770
PerCP-Vio 700
Vio Bright B515
VioBlue
VioGreen

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:

130-116-748, 130-116-627

Protocols:

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

Description:

Peripheral blood mononuclear cells (PBMCs) from rhesus monkeys were stained with CD3 antibodies or with the corresponding REA Control (S) antibodies (left images) as well as with CD45 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 or 4',6-diamidino-2-phenylindole (DAPI) fluorescence, as in the case of tandem conjugates.

Extended validation for CD3 Antibody, anti-non-human primate, 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 REA994
FN-18	++
10D12	++
Cells were incubated with an excess of purified unconjugated CD3 (REA994) 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 CD3. Rhesus monkey peripheral blood mononuclear cells (PBMCs) were stained with CD3antibodies and with a suitable counterstaining. As a control, CD3 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 CD3. Rhesus monkey peripheral blood mononuclear cells (PBMCs) were stained with CD3antibodies and with a suitable counterstaining. As a control, CD3 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 CD3. Rhesus monkey peripheral blood mononuclear cells (PBMCs) were stained with CD3antibodies and with a suitable counterstaining. As a control, CD3 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 CD3 (REA994). 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 CD3 (REA994). 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 CD3 Antibody, anti-non-human primate, REAfinity™

Overview

Clone REA994 recognizes rhesus monkey (

Macaca mulatta

) and cynomolgus monkey (

Macaca fascicularis

) CD3 T cells. The antibody has been reported to be cross-reactive with pigtail monkey (

Macaca nemestrina

), olive baboon monkey (

Papio anubis

), and african green monkey (

Chlorocebus aethiops

) T cells. CD3 is expressed on all T cells and on a subset of NK cells. It is associated with the T cell receptor (TCR) and is responsible for its signal transduction.

Additional information: Clone REA994 displays negligible binding to Fc receptors.

Alternative names

T3

Detailed product information

Technical specifications

Clone	REA994
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	non-human primate
Cross-reactivity	pigtail monkey ( Macaca nemestrina ) , olive baboon ( Papio anubis ) , african green monkey ( Chlorocebus aethiops )
Antigen	CD3
Alternative names of antigen	T3
Distribution of antigen	T cells, NK cells
Entrez Gene ID	699582, 705270, 697814, 699467
RRID	AB_2727629, AB_2727675, AB_2727630, AB_2727676, AB_2727631, AB_2727680, AB_2727636, AB_2727681, AB_2727637, AB_2727682, AB_2727638, AB_2727677, AB_2727632, AB_2727633, AB_2727678, AB_2727634, AB_2727679, AB_2727635, AB_2727673, AB_2727628, AB_2727674

Resources for CD3 Antibody, anti-non-human primate, REAfinity™

Certificates

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Applications

Extended validation