Clone:
RA3-6B2
Type of antibody:
Primary antibodies
Isotype:
rat IgG2aκ
Applications:
FC, MICS, IF, IHC
Alternative names:
Ptprc, B220, CD45, loc, L-CA, Ly-5, Lyt-4, T200

Extended validation for CD45R (B220) Antibody, anti-mouse

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 RA3-6B2
REA755++
Cells were incubated with an excess of purified unconjugated CD45R (B220) (RA3-6B2) 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 CD45R (B220). Splenocytes from C57BL/6 mice were stained with CD45R (B220) antibodies and with a suitable counterstaining. As a control, CD45R (B220) 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 CD45R (B220). Splenocytes from C57BL/6 mice were stained with CD45R (B220) antibodies and with a suitable counterstaining. As a control, CD45R (B220) 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 CD45R (B220). Splenocytes from C57BL/6 mice were stained with CD45R (B220) antibodies and with a suitable counterstaining. As a control, CD45R (B220) 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 CD45R (B220). Splenocytes from C57BL/6 mice were stained with CD45R (B220) antibodies and with a suitable counterstaining. As a control, CD45R (B220) 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 (B220). Splenocytes from C57BL/6 mice were stained with CD45R (B220) antibodies and with a suitable counterstaining. As a control, CD45R (B220) 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 CD45R (B220) (RA3-6B2). 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 CD45R (B220) (RA3-6B2). 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 (B220) (RA3-6B2). 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 (B220) Antibody, anti-mouse

Overview

Clone RA3-6B2 recognizes the mouse CD45R (B220) antigen 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. Apart from B cells, CD45R is expressed on a small subset of dendritic cells (plasmacytoid dendritic cells). The CD45R monoclonal antibody clone RA3-6B2 specifically recognizes the exon A–restricted isoform of mouse CD45. CD45R is absent in thymus but reported to be present on apoptotic thymocytes.

Alternative names

Ptprc, B220, CD45, loc, L-CA, Ly-5, Lyt-4, T200

Detailed product information

Technical specifications

CloneRA3-6B2
Clonalitymonoclonal
Isotyperat IgG2aκ
Isotype controlIsotype Control Antibody, rat IgG2a
Hostrat
Type of antibodyPrimary antibodies
Speciesmouse
AntigenCD45R (B220)
Alternative names of antigenPtprc, B220, CD45, loc, L-CA, Ly-5, Lyt-4, T200
Molecular mass of antigen [kDa]142
Distribution of antigenB cells, NK cells, T cells
Entrez Gene ID19264
RRIDAB_2751482, AB_2733944, AB_2733945, AB_2734088, AB_2734089, AB_2733914, AB_2733915, AB_2752023, AB_2751992, AB_2819495, AB_2811529, AB_2819526, AB_2876916, AB_2660450, AB_2660451, AB_2660452, AB_2751518

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