Clone:
HB5
Isotype:
mouse IgG2aκ
Applications:
MC, FC, MICS, IF, IHC
Alternative names:
CR2, C3dR, CR, CVID7, SLEB9
Type of antibody:
Primary antibodies

Extended validation for CD21 Antibody, anti-human

Specificity

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
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Fluorescence microscopy image of CD21 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD21-PE (HB5, red) and counterstained with DRAQ5 (blue) as DNA stain.
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Fluorescence microscopy image of CD21 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD21-PE (HB5, 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 (HB5, red) and counterstained with DRAQ5 (blue) as DNA stain.
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Overlay histogram showing flow cytometric analysis of CD21 knockout cells. Wild type (red) and knockout cells (blue) were stained with CD21-PE, clone (HB5). 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 (HB5). 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.
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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.
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
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Comparison of staining pattern on non-fixed and fixed cells using CD21 (HB5). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
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Comparison of staining pattern on non-fixed and fixed cells using CD21 (HB5). 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 CD21 (HB5). 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 CD21 Antibody, anti-human

Overview

Clone HB5 recognizes CD21, 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.

Alternative names

CR2, C3dR, CR, CVID7, SLEB9

Detailed product information

Technical specifications

CloneHB5
Clonalitymonoclonal
Isotypemouse IgG2aκ
Isotype controlIsotype Control Antibody, mouse IgG2a
Hostmouse
Type of antibodyPrimary antibodies
Specieshuman, non-human primate
Cross-reactivity
common marmoset (
Callithrix jacchus
)
AntigenCD21
Alternative names of antigenCR2, C3dR, CR, CVID7, SLEB9
Distribution of antigenB cells, dendritic cells, T cells, T cells, B cells, dendritic cells
RRIDAB_2656278, AB_2656279, AB_2656280, AB_2656281, AB_2656282, AB_2656283, AB_2656284, AB_2656285, AB_2656286, AB_2656287, AB_2656288, AB_2656289, AB_2656290, AB_2656291, AB_2656292, AB_2656293, AB_2656294

References for CD21 Antibody, anti-human

Publications

  1. Barrington, R. A. et al. (2005) CD21/CD19 coreceptor signaling promotes B cell survival during primary immune responses. J. Immunol. 175: 2589-2867
  2. 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
  3. 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
  4. 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

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