CD45R Antibody, anti-rat, REAfinity™

Name: CD45R Antibody, anti-rat, REAfinity™
Availability: InStock

CD45R Antibody, anti-rat, REAfinity™Extended ValidationRecombinant

Clone:

REA450

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

FC

Alternative names:

Ptprc, T200, L-CA, LCA, RT7, CD45

Flow cytometry applications forCD45R Antibody, anti-rat, REAfinity™

APC
APC-Vio 770
Biotin
PE
PE-Vio 770
Vio Bright FITC

Conjugate:

APC

Recommended dilution:

1:10

Remark:

Cells should be stained prior to fixation, if formaldehyde is used as a fixative.

Tested:

QC tested

Products used:

130-106-827, 130-106-775

Protocols:

Cell surface flow cytometry staining protocol (PBS/EDTA/BSA 1:10)

Description:

Splenocytes from Wistar rats were stained with CD45R antibodies or with the corresponding REA Control antibodies (left images) and analyzed by flow cytometry using the MACSQuant^® Analyzer. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.

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:

130-128-081

Protocols:

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

Description:

Splenocytes from Wistar rats were stained with CD45R antibodies or with the corresponding REA Control antibodies (left images) and analyzed by flow cytometry using the MACSQuant^® Analyzer. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.

Extended validation for CD45R 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 REA450
HIS24	++
Cells were incubated with an excess of purified unconjugated CD45R (REA450) 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 CD45R. Splenocytes from Wistar rat were stained with CD45R antibodies and with a suitable counterstaining. As a control, CD45R 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 CD45R. Splenocytes from Wistar rat were stained with CD45R antibodies and with a suitable counterstaining. As a control, CD45R 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 CD45R. Splenocytes from Wistar rat were stained with CD45R antibodies and with a suitable counterstaining. As a control, CD45R 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 CD45R (REA450). 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 CD45R (REA450). 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 CD45R Antibody, anti-rat, REAfinity™

Overview

Clone REA450 recognizes the rat CD45R antigen, an molecule which is expressed on B lymphocytes throughout their development from early pro–B stages onwards and is down-regulated upon terminal differentiation to plasma cells. Expression levels of CD45R are different among distinct B cell subpopulations and appears to be an indicator of maturational stages.

Additional information: Clone REA450 displays negligible binding to Fc receptors.

Alternative names

Ptprc, T200, L-CA, LCA, RT7, CD45

Detailed product information

Technical specifications

Clone	REA450
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	CD45R
Alternative names of antigen	Ptprc, T200, L-CA, LCA, RT7, CD45
Molecular mass of antigen [kDa]	141
Distribution of antigen	B cells, lymphocytes, plasma cells
Entrez Gene ID	24699
RRID	AB_2784213, AB_2658294, AB_2658295, AB_2658296, AB_2658297, AB_2658302, AB_2658303, AB_2905027, AB_2905028, AB_2922099, AB_2784214

Resources for CD45R Antibody, anti-rat, REAfinity™

Certificates

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References for CD45R Antibody, anti-rat, REAfinity™

Publications

Opstelten, D. et al. (1986) B lymphocyte-associated antigens on terminal deoxynucleotidyl transferase-positive cells and pre-B cells in bone marrow of the rat. J Immunol 137(1): 76-84
Kroese, F. G. et al. (1987) B lymphocyte differentiation in the rat: production and characterization of monoclonal antibodies to B lineage-associated antigens. Eur. J. Immunol. 17(7): 921-928
Kroese, F. G. et al. (1990) The rat B cell system: the anatomical localization of flow cytometry-defined B cell subpopulations. Eur. J. Immunol. 20(7): 1527-1534

Applications

Extended validation