CD4 (Domain 1) Antibody, anti-rat, REAfinity™

Name: CD4 (Domain 1) Antibody, anti-rat, REAfinity™
Availability: InStock

CD4 (Domain 1) Antibody, anti-rat, REAfinity™Extended ValidationRecombinant

Clone:

REA482

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

FC

Alternative names:

W3/25, p55, Leu-3

Flow cytometry applications forCD4 (Domain 1) Antibody, anti-rat, REAfinity™

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

130-126-109

Protocols:

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

Description:

Splenocytes from Wistar rats were stained with CD4 (Domain 1) antibodies or with the corresponding REA Control antibodies (left image) as well as with CD3 antibodies. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.

Extended validation for CD4 (Domain 1) Antibody, anti-rat, 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 REA482
OX-38	++
OX-35	-
W3/25	++
REA489	-
Cells were incubated with an excess of purified unconjugated CD4 (Domain 1) (REA482) 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 (Domain 1). Splenocytes from Wistar rat were stained with CD4 (Domain 1) antibodies and with a suitable counterstaining. As a control, CD4 (Domain 1) 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 CD4 (Domain 1). Splenocytes from Wistar rat were stained with CD4 (Domain 1) antibodies and with a suitable counterstaining. As a control, CD4 (Domain 1) 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 CD4 (Domain 1). Splenocytes from Wistar rat were stained with CD4 (Domain 1) antibodies and with a suitable counterstaining. As a control, CD4 (Domain 1) 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 CD4 (Domain 1). Splenocytes from Wistar rat were stained with CD4 (Domain 1) antibodies and with a suitable counterstaining. As a control, CD4 (Domain 1) 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 CD4 (Domain 1) (REA482). 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 (Domain 1) (REA482). 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 (Domain 1) Antibody, anti-rat, REAfinity™

Overview

Clone REA482 recognizes domain 1 of the rat CD4 antigen, a single-pass type I membrane protein also known as T cell surface antigen T4/Leu-3 or W3/25 antigen. CD4 is a member of the immunoglobulin superfamily and is expressed on the surface of T helper cells, monocytes, macrophages, and dendritic cells. It has four immunoglobulin domains (D1 to D4). D1 and D3 resemble immunoglobulin variable (IgV) domains, and D2 and D4 resemble immunoglobulin constant (IgC) domains. CD4 is a co-receptor that assists the T cell receptor (TCR) in communicating with an antigen-presenting cell (APC). CD4 amplifies the signal generated by the TCR by recruiting the tyrosine kinase Lck, which is essential for activating many molecular components of the signaling cascade of an activated T cell. CD4 also interacts directly with MHC class II molecules on the surface of APCs.

Additional information: Clone REA482 displays negligible binding to Fc receptors.

Alternative names

W3/25, p55, Leu-3

Detailed product information

Technical specifications

Clone	REA482
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	rat
Antigen	CD4 (Domain 1)
Alternative names of antigen	W3/25, p55, Leu-3
Molecular mass of antigen [kDa]	48
Distribution of antigen	dendritic cells, macrophages, monocytes, T cells, thymocytes, T helper cells
Entrez Gene ID	24932
RRID	AB_2857579, AB_2857759, AB_2657927, AB_2657928, AB_2657929, AB_2657930, AB_2657937, AB_2657938, AB_2657939, AB_2657940, AB_2921897, AB_2857838

Resources for CD4 (Domain 1) Antibody, anti-rat, REAfinity™

Certificates

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Reviews for CD4 (Domain 1) Antibody, anti-rat, REAfinity™

Good Separation of CD4 Population

1
2
3
4
5

CD4 (Domain 1)-PE, rat (130-107-668)

we analyzied CD 4 and CD8 population on lymph nodes . We chose milteny antibodies and got good results.

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References for CD4 (Domain 1) Antibody, anti-rat, REAfinity™

Publications

Clark, S. J. et al. (1987) Peptide and nucleotide sequences of rat CD4 (W3/25) antigen: evidence for derivation from a structure with four immunoglobulin-related domains. Proc. Natl. Acad. Sci. U.S.A. 84(6): 1649-1653
Jin, H. et al. (2013)
CD4
⁺
CD25
⁺
Foxp3
⁺
T cells contribute to the antiasthmatic effects of
Astragalus membranaceus
extract in a rat model of asthma.
Int. Immunopharmacol. 15(1): 42-49
Almolda, B. et al. (2009) CD4 microglial expression correlates with spontaneous clinical improvement in the acute Lewis rat EAE model. J. Neuroimmunol. 209(1-2): 65-80

Applications

Extended validation