Clone:
REA780
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC, MC
Alternative names:
MS4A1, B1, Bp35, CVID5, LEU-16, MS4A2, S7, Ly-44

Extended validation for CD20 Antibody, anti-human, 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 REA780
LT20++
2H7++
Hu2++
LT27++
Cells were incubated with an excess of purified unconjugated CD20 (REA780) 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. Human peripheral blood mononuclear cells (PBMCs) 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. Human peripheral blood mononuclear cells (PBMCs) 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. Human peripheral blood mononuclear cells (PBMCs) 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. Human peripheral blood mononuclear cells (PBMCs) 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. Human peripheral blood mononuclear cells (PBMCs) 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 (REA780). 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 (REA780). 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 (REA780). 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-human, REAfinity™

Overview

Clone REA780 recognizes the CD20 antigen, a non-glycosylated transmembrane protein of 33–37 kDa that is expressed on B lineage cells from the pre–B cell stage to the B cell lymphoblast stage. The antigen is further expressed on most malignant B cells. CD20 is not found on early B cell progenitors or plasma cells. Oligomers of CD20 form a Ca
2+
channel and might function in the regulation of local responses during B cell activation.
Additional information: Clone REA780 displays negligible binding to Fc receptors.

Alternative names

MS4A1, B1, Bp35, CVID5, LEU-16, MS4A2, S7, Ly-44

Detailed product information

Technical specifications

CloneREA780
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
Cross-reactivity
cynomolgus monkey (
Macaca fascicularis
)
,
rhesus monkey (
Macaca mulatta
)
AntigenCD20
Alternative names of antigenMS4A1, B1, Bp35, CVID5, LEU-16, MS4A2, S7, Ly-44
Molecular mass of antigen [kDa]33
Distribution of antigenB cells, dendritic cells, lymphocytes, cancer stem cells
Entrez Gene ID931
RRIDAB_2656062, AB_2656063, AB_2656064, AB_2656065, AB_2656066, AB_2656067, AB_2656068, AB_2656071, AB_2656072, AB_2656073, AB_2656074, AB_2656075, AB_2656076, AB_2656077, AB_2656078, AB_2656079, AB_2656080, AB_2656081, AB_2656082, AB_2656061

Resources for CD20 Antibody, anti-human, REAfinity™

Certificates

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

References for CD20 Antibody, anti-human, REAfinity™

Publications

  1. Countouriotis, T. B. et al. (2002) Cell surface antigen and molecular targeting in the treatment of hematologic malignancies. Stem Cells 20(3): 215-229
  2. Polyak et al. (2002) CD20 Workshop Panel report. In: Leucocyte typing VII, Oxford University Press.
  3. Lamprecht, B. et al. (2008) Aberrant expression of the Tʜ2 cytokine IL-21 in Hodgkin lymphoma cells regulates STAT3 signaling and attracts Treg cells via regulation of MIP-3alpha. Blood 112(8): 3339-3347
  4. Maxwell, S. A. et al. (2009) 14-3-3zeta J. Biol. Chem. 284(33): 22379-22389

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