Clone:
REA482
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC
Alternative names:
W3/25, p55, Leu-3

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 clonesOverlap 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.
View details
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.
View details
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.
View details
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.
View details
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
View details
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.
View details
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

CloneREA482
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody, human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Speciesrat
AntigenCD4 (Domain 1)
Alternative names of antigenW3/25, p55, Leu-3
Molecular mass of antigen [kDa]48
Distribution of antigendendritic cells, macrophages, monocytes, T cells, thymocytes, T helper cells
Entrez Gene ID24932
RRIDAB_2857579, AB_2857759, AB_2657927, AB_2657928, AB_2657929, AB_2657930, AB_2657937, AB_2657938, AB_2657939, AB_2657940, AB_2857838

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.

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

Publications

  1. 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
  2. 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
  3. 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

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