Clone:
REA737
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC, 3D-IF
Alternative names:
L-CA, LCA, Ly-5, T200

Extended validation for CD45 Antibody, anti-mouse, 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 REA737
30F11++
I3/2.3+
IBL-5/25++
Cells were incubated with an excess of purified unconjugated CD45 (REA737) 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 CD45. Splenocytes from C57BL/6 mice were stained with CD45 antibodies and plotted against the side scatter. As a control, CD45 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 CD45. Splenocytes from C57BL/6 mice were stained with CD45 antibodies and plotted against the side scatter. As a control, CD45 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 CD45. Splenocytes from C57BL/6 mice were stained with CD45 antibodies and plotted against the side scatter. As a control, CD45 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 CD45. Splenocytes from C57BL/6 mice were stained with CD45 antibodies and plotted against the side scatter. As a control, CD45 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 CD45. Splenocytes from C57BL/6 mice were stained with CD45 antibodies and plotted against the side scatter. As a control, CD45 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 CD45. Splenocytes from C57BL/6 mice were stained with CD45 antibodies and plotted against the side scatter. As a control, CD45 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 CD45. Splenocytes from C57BL/6 mice were stained with CD45 antibodies and plotted against the side scatter. As a control, CD45 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 CD45 (REA737). 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 CD45 (REA737). 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 CD45 (REA737). 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 CD45 Antibody, anti-mouse, REAfinity™

Overview

Clone REA737 recognizes the mouse CD45 antigen, also known as Ly-5 or leukocyte common antigen (LCA), which belongs to the protein tyrosine phosphatase (PTP) family. CD45 is a glycoprotein which is involved in T cell receptor and B cell receptor signal transduction. It is expressed at high levels on all cells of hematopoietic origin except for erythrocytes. Clone REA737 reacts with all CD45 isoforms. CD45 can be used to discriminate leukocytes from non-hematopoietic cells.
Additional information: Clone REA737 displays negligible binding to Fc receptors.

Alternative names

L-CA, LCA, Ly-5, T200

Detailed product information

Technical specifications

CloneREA737
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody, human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Speciesmouse
AntigenCD45
Alternative names of antigenL-CA, LCA, Ly-5, T200
Molecular mass of antigen [kDa]142
Distribution of antigenT cells, B cells
Entrez Gene ID19264
RRIDAB_2658217, AB_2658218, AB_2658219, AB_2658220, AB_2658221, AB_2658222, AB_2658223, AB_2658224, AB_2658225, AB_2658226, AB_2658227, AB_2658228, AB_2658229, AB_2658230, AB_2658231, AB_2658232, AB_2658233, AB_2658234, AB_2658235, AB_2658216

References for CD45 Antibody, anti-mouse, REAfinity™

Publications

  1. Pulido, R. et al. (1988) Comparative biochemical and tissue distribution study of four distinct CD45 antigen specificities. J. Immunol. 140(11): 3851-3857
  2. Arendt, C. W. et al. (1997) Identification of the CD45-associated 116 kDa and 80 kDa proteins as the alpha- and beta-subunits of alpha-glucosidase II. J. Biol. Chem. 272(20): 13117-13125
  3. Trowbridge, I. S. et al. (1994) CD45: an emerging role as a protein tyrosine phosphatase required for lymphocyte activation and development. Annu. Rev. Immunol. 12: 85-116

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