Clone:
REA1047
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC
Alternative names:
Ptprc, B220, CD45, CD45R, GP180, L-CA, Ly-5, T200

Extended validation for CD45RA Antibody, anti-human, 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 REA1047
HI100++
JS-83-
L48-
T6D11-
REA562-
Cells were incubated with an excess of purified unconjugated CD45RA (REA1047) 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.
Knockout validation
To ensure antibody specificity, the target gene is knocked out in a suitable cell line using the CRISPR/Cas9 system and the knockout is confirmed by sequencing of the target locus. The antibody is considered to bind specifically to the intended epitope if no antibody binding to the knockout cells can be detected. The antibody staining is controlled by fluorescence microscopy and/or flow cytometry.
WT
KO
View details
Fluorescence microscopy image of CD45RA knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD45RA-PE (REA1047, red) and counterstained with DRAQ5 (blue) as DNA stain.
View details
Fluorescence microscopy image of CD45RA knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD45RA-PE (REA1047, red) and counterstained with DRAQ5 (blue) as DNA stain.
Fluorescence microscopy image of CD45RA knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD45RA-PE (REA1047, red) and counterstained with DRAQ5 (blue) as DNA stain.
View details
Overlay histogram showing flow cytometric analysis of CD45RA knockout cells. Wild type (red) and knockout cells (blue) were stained with CD45RA-PE, clone (REA1047). 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 propidium iodide fluorescence.
Overlay histogram showing flow cytometric analysis of CD45RA knockout cells. Wild type (red) and knockout cells (blue) were stained with CD45RA-PE, clone (REA1047). 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 propidium iodide fluorescence.

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 CD45RA. Human peripheral blood mononuclear cells (PBMCs) were stained with CD45RA antibodies and with a suitable counterstaining. As a control, CD45RA 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 CD45RA. Human peripheral blood mononuclear cells (PBMCs) were stained with CD45RA antibodies and with a suitable counterstaining. As a control, CD45RA 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 CD45RA. Human peripheral blood mononuclear cells (PBMCs) were stained with CD45RA antibodies and with a suitable counterstaining. As a control, CD45RA 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 CD45RA. Human peripheral blood mononuclear cells (PBMCs) were stained with CD45RA antibodies and with a suitable counterstaining. As a control, CD45RA 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 CD45RA. Human peripheral blood mononuclear cells (PBMCs) were stained with CD45RA antibodies and with a suitable counterstaining. As a control, CD45RA 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 CD45RA. Human peripheral blood mononuclear cells (PBMCs) were stained with CD45RA antibodies and with a suitable counterstaining. As a control, CD45RA 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 CD45RA. Human peripheral blood mononuclear cells (PBMCs) were stained with CD45RA antibodies and with a suitable counterstaining. As a control, CD45RA 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 CD45RA (REA1047). 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 CD45RA (REA1047). 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 CD45RA (REA1047). 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 CD45RA Antibody, anti-human, REAfinity™

Overview

Clone REA1047 recognizes the human CD45RA antigen which is expressed on naive CD4
+
and CD8
+
T cells as well as on CD8
+
effector T cells. CD45RA is also present on subsets of B and NK cells as well as on plasmacytoid dendritic cells. The CD45RA antibody recognizes the 220 kDa isoform of the leukocyte common antigen (LCA), a transmembrane tyrosine phosphatase.
Additional information: Clone REA1047 displays negligible binding to Fc receptors.

Alternative names

Ptprc, B220, CD45, CD45R, GP180, L-CA, Ly-5, T200

Detailed product information

Technical specifications

CloneREA1047
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman, non-human primate
Cross-reactivity
chimpanzee (
Pan troglodytes
)
AntigenCD45RA
Alternative names of antigenPtprc, B220, CD45, CD45R, GP180, L-CA, Ly-5, T200
Molecular mass of antigen [kDa]145
Distribution of antigenB cells, monocytes, NK cells, T cells, CD4+ T cells, CD8+ T cells, naive T cells, thymocytes
Entrez Gene ID5788
RRIDAB_2733642, AB_2733814, AB_2733815, AB_2733376, AB_2733377, AB_2733946, AB_2733947, AB_2734090, AB_2734091, AB_2732976, AB_2732977, AB_2733236, AB_2733237, AB_2733125, AB_2733126, AB_2733033, AB_2733034, AB_2734021, AB_2734022, AB_2733524, AB_2733525, AB_2733641

Resources for CD45RA Antibody, anti-human, REAfinity™

Certificates

Please follow this
link
to search for Certificates of Analysis (CoA) by lot number.

References for CD45RA Antibody, anti-human, REAfinity™

Publications

  1. Cantisan, S. et al. (2010)
    CD45RA expression on HCMV-specific effector memory CD8
    +
    T cells is associated with the duration and intensity of HCMV replication after transplantation.
    Clin. Immunol. 137(1): 81-88
  2. Braun, T. et al. (2012)
    Effector CD4
    +
    CD45RA
    CD25
    bright
    Foxp3
    bright
    regulatory T cell (eTreg) distribution is significantly impaired in chronic myelomonocytic leukemia (CMML) and correlates with TET 2 mutational status.
    Blood 120: 2808-2808

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