Clone:
REA504
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC
Alternative names:
Ptprc, L-CA, T200

Extended validation for CD45 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 REA504
OX-1++
Cells were incubated with an excess of purified unconjugated CD45 (REA504) 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 Wistar rat were stained with CD45 antibodies and with a suitable counterstaining. 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 Wistar rat were stained with CD45 antibodies and with a suitable counterstaining. 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 Wistar rat were stained with CD45 antibodies and with a suitable counterstaining. 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 Wistar rat were stained with CD45 antibodies and with a suitable counterstaining. 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 (REA504). 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 (REA504). 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 (REA504). 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-rat, REAfinity™

Overview

Clone REA504 recognizes the rat CD45 antigen, also known as leukocyte common antigen (LCA), which is a 180–220 kDa type I transmembrane protein with multiple isoforms and differential glycosylation. It appears in at least five isoforms depending on the differentiation status of the cell. CD45 is expressed at high levels on all cells of hematopoietic origin except for erythrocyte and platelets. It plays a role in signal transduction through B and T cell antigen receptors.
Additional information: Clone REA504 displays negligible binding to Fc receptors.

Alternative names

Ptprc, L-CA, T200

Detailed product information

Technical specifications

CloneREA504
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody, human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Speciesrat
AntigenCD45
Alternative names of antigenPtprc, L-CA, T200
Molecular mass of antigen [kDa]141
Distribution of antigenleukocytes
Entrez Gene ID24699
RRIDAB_2752034, AB_2751998, AB_2819438, AB_2857718, AB_2658258, AB_2658259, AB_2658260, AB_2658261, AB_2658262, AB_2658263

Resources for CD45 Antibody, anti-rat, REAfinity™

References for CD45 Antibody, anti-rat, REAfinity™

Publications

  1. Sunderland, C. A. et al. (1979) Purification with monoclonal antibody of a predominant leukocyte-common antigen and glycoprotein from rat thymocytes. Eur. J. Immunol. 9(2): 155-159
  2. Mojcik, D. L. et al. (1987) Monoclonal antibodies to RT7 and LCA antigens in the rat: cell distribution and segregation analysis. Hybridoma 6(5): 531-543

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