Clone:
REA755
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
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, 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 REA755
RA3-6B2++
Cells were incubated with an excess of purified unconjugated CD45R (B220) (REA755) 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) (REA755). 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) (REA755). 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) (REA755). 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, REAfinity™

Overview

Clone REA755 recognizes the mouse CD45R (B220) antigen, a B lineage-specific surface 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. Apart from B cells, CD45R is expressed on a small subset of dendritic cells (plasmacytoid dendritic cells). CD45R is absent in thymus but reported to be present on apoptotic thymocytes. Clone REA755 specifically recognizes the exon A–restricted isoform of mouse CD45.
Additional information: Clone REA755 displays negligible binding to Fc receptors.

Alternative names

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

Detailed product information

Technical specifications

CloneREA755
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody, human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Speciesmouse
Cross-reactivityhuman
AntigenCD45R (B220)
Alternative names of antigenPtprc, B220, CD45, loc, L-CA, Ly-5, Lyt-4, T200
Molecular mass of antigen [kDa]142
Distribution of antigenT cells, B cells, NK cells
Entrez Gene ID19264
RRIDAB_2658274, AB_2658275, AB_2658276, AB_2658277, AB_2658278, AB_2658279, AB_2658280, AB_2658281, AB_2658282, AB_2658283, AB_2658284, AB_2658285, AB_2658286, AB_2658287, AB_2658288, AB_2658289, AB_2658290, AB_2658291, AB_2658273

Resources for CD45R (B220) Antibody, anti-mouse, REAfinity™

Certificates

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

References for CD45R (B220) Antibody, anti-mouse, REAfinity™

Publications

  1. Coffman, R. L. (1982) Surface antigen expression and immunoglobulin gene rearrangement during mouse pre-B cell development. Immunol. Rev. 69: 5-23
  2. Katavic, V. et al. (2003) The surface antigen CD45R identifies a population of estrogen-regulated murine marrow cells that contain osteoclast precursors. Bone 32(6): 581-590

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