Clone:
REA940
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MC
Alternative names:
CR2, C3dR, CR, CVID7, SLEB9

Extended validation for CD21 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 REA940
B-ly4-
Bu32-
HB5-
LT21++
Cells were incubated with an excess of purified unconjugated CD21 (REA940) 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.
Knockout validation
To ensure antibody specificity, the target gene is knocked out in a suitable cell line using the CRISPR/Cas9 system and the knockout is confirmed by sequencing of the target locus. The antibody is considered to bind specifically to the intended epitope if no antibody binding to the knockout cells can be detected. The antibody staining is controlled by fluorescence microscopy and/or flow cytometry.
WT
KO
View details
Fluorescence microscopy image of CD21 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD21-PE (REA940, red) and counterstained with DRAQ5 (blue) as DNA stain.
View details
Fluorescence microscopy image of CD21 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD21-PE (REA940, red) and counterstained with DRAQ5 (blue) as DNA stain.
Fluorescence microscopy image of CD21 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD21-PE (REA940, red) and counterstained with DRAQ5 (blue) as DNA stain.
View details
Overlay histogram showing flow cytometric analysis of CD21 knockout cells. Wild type (red) and knockout cells (blue) were stained with CD21-PE, clone (REA940). 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 propidium iodide fluorescence.
Overlay histogram showing flow cytometric analysis of CD21 knockout cells. Wild type (red) and knockout cells (blue) were stained with CD21-PE, clone (REA940). 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 propidium iodide fluorescence.

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 CD21. Human peripheral blood mononuclear cells (PBMCs) were stained with CD21 antibodies and with a suitable counterstaining. As a control, CD21 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 CD21. Human peripheral blood mononuclear cells (PBMCs) were stained with CD21 antibodies and with a suitable counterstaining. As a control, CD21 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 CD21. Human peripheral blood mononuclear cells (PBMCs) were stained with CD21 antibodies and with a suitable counterstaining. As a control, CD21 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 CD21. Human peripheral blood mononuclear cells (PBMCs) were stained with CD21 antibodies and with a suitable counterstaining. As a control, CD21 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 CD21. Human peripheral blood mononuclear cells (PBMCs) were stained with CD21 antibodies and with a suitable counterstaining. As a control, CD21 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 CD21. Human peripheral blood mononuclear cells (PBMCs) were stained with CD21 antibodies and with a suitable counterstaining. As a control, CD21 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 CD21. Human peripheral blood mononuclear cells (PBMCs) were stained with CD21 antibodies and with a suitable counterstaining. As a control, CD21 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 CD21. Human peripheral blood mononuclear cells (PBMCs) were stained with CD21 antibodies and with a suitable counterstaining. As a control, CD21 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.

Specifications for CD21 Antibody, anti-human, REAfinity™

Overview

Clone REA940 recognizes the human CD21 antigen, a type I membrane glycoprotein. Expression of CD21 in humans is found on B cells, follicular dendritic cells, subsets of epithelial cells, and thymic T cells. The primary function attributed to CD21 has been to amplify the B cell receptor (BCR)–mediated signal transduction in response to antigen recognition. To enhance the BCR mediated activation, CD21 associates with CD19 and CD81 in a B cell–specific signal transduction complex, where interaction of BCR with CR2/CD19 amplifies the signals. In addition, CD21 serves as a receptor for split products of complement protein C3, the gp350/220 viral coat protein of the EBV and the immunoregulatory protein CD23.
Additional information: Clone REA940 displays negligible binding to Fc receptors.

Alternative names

CR2, C3dR, CR, CVID7, SLEB9

Detailed product information

Technical specifications

CloneREA940
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman, other
Cross-reactivitycow, dog, pig
AntigenCD21
Alternative names of antigenCR2, C3dR, CR, CVID7, SLEB9
Molecular mass of antigen [kDa]111
Distribution of antigenB cells, dendritic cells, T cells
Entrez Gene ID1380
RRIDAB_2727066, AB_2727122, AB_2727067, AB_2727123, AB_2727068, AB_2727124, AB_2727069, AB_2727120, AB_2727065, AB_2811646, AB_2727121

References for CD21 Antibody, anti-human, REAfinity™

Publications

  1. Roberts, M. L. et al. (1996) Epstein—Barr virus binding to CD21, the virus receptor, activates resting B cells via an intracellular pathway that is linked to B cell infection. Virology 77: 3077-3085
  2. Braun, M. et al. (1998) Human B and T lymphocytes have similar amounts of CD21 mRNA, but differ in surface expression of the CD21 glycoprotein. Int. Immunol. 10: 1197-1202
  3. Cherukuri, A. et al. (2001) The role of the CD19/CD21 complex in B cell processing and presentation of complement-tagged antigens. J. Immunol. 167: 163-172
  4. Barrington, R. A. et al. (2005) CD21/CD19 coreceptor signaling promotes B cell survival during primary immune responses. J. Immunol. 175: 2589-2867

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