CD3 Antibody, anti-human

Name: CD3 Antibody, anti-human
Availability: InStock

CD3 Antibody, anti-humanExtended Validation

Clone:

BW264/56

Type of antibody:

Primary antibodies

Isotype:

mouse IgG2aκ

Applications:

FC, MICS, IF, IHC, ICC

Alternative names:

CD3e, IMD18, T3E, TCRE

Flow cytometry applications forCD3 Antibody, anti-human

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

Reported:

23289765, 24518760, 28400477, all publications

Products used:

130-113-687, 130-113-125

Protocols:

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

Description:

Human peripheral blood mononuclear cells (PBMCs) were stained with CD3 antibodies and analyzed by flow cytometry 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.

MICS (MACSima Imaging Cyclic Staining) applications forCD3 Antibody, anti-human

APC
FITC
PE

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

PFA

Tested:

during development

Species tested:

human

Products used:

130-113-687, 130-113-125

Compatible applications:

IF, IHC, ICC

Protocols:

Protocol for MICS experiments

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

Acetone

Tested:

during development

Species tested:

human

Products used:

130-113-687, 130-113-125

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Extended validation for CD3 Antibody, anti-human

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 recognition with BW264/56
REA613	++
UCHT1	++
HIT3	++
SK7	++
OKT3	++
SP34-2	+
Cells were incubated with an excess of purified unconjugated CD3 (BW264/56) 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. Human peripheral blood mononuclear cells (PBMCs) were stained with CD3 antibodies 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. Human peripheral blood mononuclear cells (PBMCs) were stained with CD3 antibodies 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. Human peripheral blood mononuclear cells (PBMCs) were stained with CD3 antibodies 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. Human peripheral blood mononuclear cells (PBMCs) were stained with CD3 antibodies 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. Human peripheral blood mononuclear cells (PBMCs) were stained with CD3 antibodies 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. Human peripheral blood mononuclear cells (PBMCs) were stained with CD3 antibodies 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. Human peripheral blood mononuclear cells (PBMCs) were stained with CD3 antibodies 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. Human peripheral blood mononuclear cells (PBMCs) were stained with CD3 antibodies 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. Human peripheral blood mononuclear cells (PBMCs) were stained with CD3 antibodies 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 (BW264/56). 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 (BW264/56). 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-human

Overview

The CD3 antibody recognizes the human CD3 antigen which is present on mature human T cells, thymocytes, and a subset of NK cells. CD3 is associated with the T cell receptor (TCR) and is responsible for its signal transduction. The CD3 antigen is a complex of five invariable chains: γ, δ, ε, ζ, and η. The epitope recognized by the antibody is located on the ε-chain of the CD3 complex.

Alternative names

CD3e, IMD18, T3E, TCRE

Detailed product information

Technical specifications

Clone	BW264/56
Clonality	monoclonal
Isotype	mouse IgG2aκ
Isotype control	Isotype Control Antibody, mouse IgG2a
Host	mouse
Type of antibody	Primary antibodies
Species	human
Antigen	CD3
Alternative names of antigen	CD3e, IMD18, T3E, TCRE
Molecular mass of antigen [kDa]	56(sum of molecular weights of subunits)
Distribution of antigen	NK cells, T cells, thymocytes
Entrez Gene ID	915, 916, 917
RRID	AB_2726236, AB_2725961, AB_2726701, AB_2726237, AB_2725962, AB_2726234, AB_2725959, AB_2726233, AB_2725958, AB_2726229, AB_2725954, AB_2726235, AB_2725960, AB_2726230, AB_2725955, AB_2725956, AB_2726232, AB_2725957, AB_2726228, AB_2725953, AB_2726231

Resources for CD3 Antibody, anti-human

Certificates

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Reviews for CD3 Antibody, anti-human

Staining Peripheral Blood with Anti-CD3 FITC (Clone BW264/56)

1
2
3
4
5

CD3-FITC, human (130-113-128)

Works well. Reproducible results.

Staining Peripheral Blood with Anti-CD3 FITC (Clone BW264/56)

1
2
3
4
5

CD3-FITC, human (130-113-690)

Works well. Reproducible results.

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References for CD3 Antibody, anti-human

Publications

Vladimir, S. et al. (2009) Human term placenta as a source of hematopoietic cells. Exp. Biol. Med. (Maywood) 234(7): 813-823

Applications

Extended validation