Clone:
REA294
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC
Alternative names:
MS4A1, Ly-44, MS4A2

Extended validation for CD20 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 REA294
SA271G2-
SA275A11-
AISB12++
Cells were incubated with an excess of purified unconjugated CD20 (REA294) 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 CD20. BALB/cAnNCrl mouse splenocytes were stained with CD20 antibodies and with a suitable counterstaining. As a control, CD20 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 CD20. BALB/cAnNCrl mouse splenocytes were stained with CD20 antibodies and with a suitable counterstaining. As a control, CD20 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 CD20. BALB/cAnNCrl mouse splenocytes were stained with CD20 antibodies and with a suitable counterstaining. As a control, CD20 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 CD20. BALB/cAnNCrl mouse splenocytes were stained with CD20 antibodies and with a suitable counterstaining. As a control, CD20 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 CD20 (REA294). 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 CD20 (REA294). 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 CD20 (REA294). 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 CD20 Antibody, anti-mouse, REAfinity™

Overview

Clone REA294 recognizes the murine CD20 antigen, a 32kDa membrane protein also known as B-cell differentiation antigen Ly-44. CD20 is a B lymphocyte-specific cell-surface molecule involved in the regulation of B-cell activation and proliferation. CD20 is expressed on B cells and a subset of T cells, but not on plasma cells.
Additional information: Clone REA294 displays negligible binding to Fc receptors.

Alternative names

MS4A1, Ly-44, MS4A2

Detailed product information

Technical specifications

CloneREA294
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody, human IgG1
Hosthuman cell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Speciesmouse
AntigenCD20
Alternative names of antigenMS4A1, Ly-44, MS4A2
Molecular mass of antigen [kDa]32
Distribution of antigenB cells
Entrez Gene ID12482
RRIDAB_2819616

Resources for CD20 Antibody, anti-mouse, REAfinity™

Certificates

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

References for CD20 Antibody, anti-mouse, REAfinity™

Publications

  1. Tedder, T. F. et al. (1988) Cloning of a complementary DNA encoding a new mouse B lymphocyte differentiation antigen, homologous to the human B1 (CD20) antigen, and localization of the gene to chromosome 19. J. Immunol. 141(12): 4388-4394
  2. Press, O. W. et al. (1987) Monoclonal antibody 1F5 (anti-CD20) serotherapy of human B cell lymphomas. Blood 69(2): 584-591
  3. Uchida, J. et al. (2004) Mouse CD20 expression and function. Int. Immunol. 16(1): 119-129

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