CD4 Antibody, anti-human

Name: CD4 Antibody, anti-human
Availability: InStock

CD4 Antibody, anti-humanExtended Validation

Clone:

M-T466

Type of antibody:

Primary antibodies

Isotype:

mouse IgG1κ

Applications:

FC

Alternative names:

CD4mut, L3T4, Leu-3, T4

Flow cytometry applications forCD4 Antibody, anti-human

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

Conjugate:

APC

Recommended dilution:

1:50

Remark:

The antibody is suited for staining of formaldehyde-fixed cells.

Tested:

QC tested

Products used:

130-113-812, 130-113-250

Protocols:

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

Description:

Human peripheral blood mononuclear cells (PBMCs) were stained with CD4(M-T466) antibodies or with the corresponding isotype control antibodies (left image) 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.

Extended validation for CD4 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 epitope recognition with M-T466
RPA-T4	-
M-A251	++
OKT4	-
VIT4	-
REA623	++
M-T321	++
Cells were incubated with an excess of purified unconjugated CD4 (M-T466) 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 CD4. Human peripheral blood mononuclear cells (PBMCs) were stained with CD4 antibodies and with a suitable counterstaining. As a control, CD4 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 CD4. Human peripheral blood mononuclear cells (PBMCs) were stained with CD4 antibodies and with a suitable counterstaining. As a control, CD4 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 CD4. Human peripheral blood mononuclear cells (PBMCs) were stained with CD4 antibodies and with a suitable counterstaining. As a control, CD4 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 CD4. Human peripheral blood mononuclear cells (PBMCs) were stained with CD4 antibodies and with a suitable counterstaining. As a control, CD4 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 CD4. Human peripheral blood mononuclear cells (PBMCs) were stained with CD4 antibodies and with a suitable counterstaining. As a control, CD4 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 CD4. Human peripheral blood mononuclear cells (PBMCs) were stained with CD4 antibodies and with a suitable counterstaining. As a control, CD4 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 CD4. Human peripheral blood mononuclear cells (PBMCs) were stained with CD4 antibodies and with a suitable counterstaining. As a control, CD4 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 CD4. Human peripheral blood mononuclear cells (PBMCs) were stained with CD4 antibodies and with a suitable counterstaining. As a control, CD4 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 CD4. Human peripheral blood mononuclear cells (PBMCs) were stained with CD4 antibodies and with a suitable counterstaining. As a control, CD4 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 CD4. Human peripheral blood mononuclear cells (PBMCs) were stained with CD4 antibodies and with a suitable counterstaining. As a control, CD4 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 CD4. Human peripheral blood mononuclear cells (PBMCs) were stained with CD4 antibodies and with a suitable counterstaining. As a control, CD4 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 CD4 (M-T466). 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 CD4 (M-T466). 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 CD4 Antibody, anti-human

Overview

CD4 is a type I transmembrane glycoprotein involved in the recognition of MHC class II/peptide complexes by the TCR heterodimers. CD4 is highly expressed on T helper cells and at a lower level on monocytes and dendritic cells. The CD4 molecule is the receptor for the human immunodeficiency virus. The CD4 antibody recognizes the human CD4 antigen which is highly expressed on human T helper cells and thymocytes, and at lower levels on monocytes and dendritic cells. It is responsible for the recognition of the MHC class II antigen.

Alternative names

CD4mut, L3T4, Leu-3, T4

Detailed product information

Technical specifications

Clone	M-T466
Clonality	monoclonal
Isotype	mouse IgG1κ
Isotype control	Isotype Control Antibody, mouse IgG1
Host	mouse
Type of antibody	Primary antibodies
Species	human, non-human primate
Cross-reactivity	rhesus monkey ( Macaca mulatta ) , cynomolgus monkey ( Macaca fascicularis )
Antigen	CD4
Alternative names of antigen	CD4mut, L3T4, Leu-3, T4
Molecular mass of antigen [kDa]	48
Distribution of antigen	dendritic cells, granulocytes, Langerhans cells, lymphocytes, macrophages, monocytes, T cells, T helper cells, thymocytes
Entrez Gene ID	920
RRID	AB_2726055, AB_2726333, AB_2726056, AB_2726329, AB_2726052, AB_2726336, AB_2726059, AB_2726337, AB_2726060, AB_2726334, AB_2726057, AB_2726330, AB_2726053, AB_2726335, AB_2726058, AB_2726331, AB_2726054, AB_2726332

Resources for CD4 Antibody, anti-human

Documents and Protocols

App notes/customer reports

Polarizing Th cells from naive CD4+ T cells

App notes/customer reports

High-sensitivity detection and analysis of rare cells by integrated magnetic enrichment and flow cytometry using the MACSQuant® Analyzer.

App notes/customer reports

Magnetic enrichment of antigen-specific CD4+ T cells using the MACSQuant® Analyzer 10 enables the in-depth characterization of vaccine-induced circulating follicular T helper cells.

App notes/customer reports

Efficient and rapid in vitro generation of fully functional multi-virus-specific CD4⁺ and CD8⁺ T cells.

App notes/customer reports

Flow cytometry analysis of Th subsets

Certificates

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link

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

References for CD4 Antibody, anti-human

Publications

Lamprecht, B. et al. (2008) Aberrant expression of the Tʜ2 cytokine IL-21 in Hodgkin lymphoma cells regulates STAT3 signaling and attracts Treg cells via regulation of MIP-3alpha. Blood 112(8): 3339-3347
Sacha, J. B. et al. (2010)
Synchronous infection of SIV and HIV
in vitro
for virology, immunology and vaccine-related studies.
Nat. Protoc. 5(2): 239-246

Applications

Extended validation