CD20 Antibody, anti-human, REAfinity™

Name: CD20 Antibody, anti-human, REAfinity™
Availability: InStock

CD20 Antibody, anti-human, REAfinity™Extended ValidationRecombinant

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

Flow cytometry applications forCD20 Antibody, anti-human, REAfinity™

APC
APC-Vio 770
Biotin
FITC
PE
PE-Vio 615
PE-Vio 770
PerCP-Vio 700
Vio Bright B515
VioBlue

Conjugate:

APC

Recommended dilution:

1:50

Remark:

The antibody is suited for staining of formaldehyde-fixed cells.

Tested:

QC tested

Products used:

130-111-525, 130-111-339

Protocols:

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

Description:

Human peripheral blood mononuclear cells (PBMCs) were stained with CD20 antibodies or with the corresponding REA Control (S) antibodies (left image) as well as with CD19 antibodies. The Tandem Signal Enhancer has been used to increase binding specificity of tandem-dye–conjugated antibodies. Flow cytometry was performed using the MACSQuant^® Analyzer. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence or 4',6-diamidino-2-phenylindole (DAPI) fluorescence, as in the case of tandem conjugates.

MICS (MACSima Imaging Cyclic Staining) applications forCD20 Antibody, anti-human, REAfinity™

APC
FITC
PE

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

PFA

Tested:

during development

Species tested:

human

Products used:

130-111-525, 130-111-339

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

Acetone

Tested:

during development

Species tested:

human

Products used:

130-111-525, 130-111-339

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Mass cytometry applications forCD20 Antibody, anti-human, REAfinity™

pure

Conjugate:

pure

Products used:

130-124-537

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 clones	Overlap 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.

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.

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

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

Clone	REA780
Clonality	monoclonal
Isotype	recombinant human IgG1
Isotype control	REA Control Antibody (S), human IgG1
Host	human cell line
Type of antibody	Primary antibodies, Recombinant antibodies
Species	human
Cross-reactivity	cynomolgus monkey ( Macaca fascicularis ) , rhesus monkey ( Macaca mulatta )
Antigen	CD20
Alternative names of antigen	MS4A1, B1, Bp35, CVID5, LEU-16, MS4A2, S7, Ly-44
Molecular mass of antigen [kDa]	33
Distribution of antigen	B cells, dendritic cells, lymphocytes, cancer stem cells
Entrez Gene ID	931
RRID	AB_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_2889505, 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

Countouriotis, T. B. et al. (2002) Cell surface antigen and molecular targeting in the treatment of hematologic malignancies. Stem Cells 20(3): 215-229
Polyak et al. (2002) CD20 Workshop Panel report. In: Leucocyte typing VII, Oxford University Press.
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
Maxwell, S. A. et al. (2009) 14-3-3zeta J. Biol. Chem. 284(33): 22379-22389

Applications

Extended validation