Clone:
REA513
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC, MC
Alternative names:
CVID6, S5.7, TAPA1, TSPAN28

Extended validation for CD81 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 REA513
JS-81++
M38++
5A6++
1D6-CD81++
Cells were incubated with an excess of purified unconjugated CD81 (REA513) 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 CD81 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD81-PE (REA513, red) and counterstained with DRAQ5 (blue) as DNA stain.
View details
Fluorescence microscopy image of CD81 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD81-PE (REA513, red) and counterstained with DRAQ5 (blue) as DNA stain.
Fluorescence microscopy image of CD81 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD81-PE (REA513, red) and counterstained with DRAQ5 (blue) as DNA stain.
View details
Overlay histogram showing flow cytometric analysis of CD81 knockout cells. Wild type (red) and knockout cells (blue) were stained with CD81-PE, clone (REAL513). 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 CD81 knockout cells. Wild type (red) and knockout cells (blue) were stained with CD81-PE, clone (REAL513). 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 CD81. Human peripheral blood mononuclear cells (PBMCs) were stained withCD81 antibodies and with a suitable counterstaining. As a control, CD81 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 CD81. Human peripheral blood mononuclear cells (PBMCs) were stained withCD81 antibodies and with a suitable counterstaining. As a control, CD81 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 CD81. Human peripheral blood mononuclear cells (PBMCs) were stained withCD81 antibodies and with a suitable counterstaining. As a control, CD81 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 CD81. Human peripheral blood mononuclear cells (PBMCs) were stained withCD81 antibodies and with a suitable counterstaining. As a control, CD81 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 CD81. Human peripheral blood mononuclear cells (PBMCs) were stained withCD81 antibodies and with a suitable counterstaining. As a control, CD81 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 CD81. Human peripheral blood mononuclear cells (PBMCs) were stained withCD81 antibodies and with a suitable counterstaining. As a control, CD81 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 CD81 (REA513). 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 CD81 (REA513). 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 CD81 (REA513). 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 CD81 Antibody, anti-human, REAfinity™

Overview

Clone REA513 recognizes the human CD81 antigen, a multi-pass membrane protein also known as TAPA-1. It is a member of the tetraspanin family including, for example, CD9, CD37, CD53, CD63, CD82, and CD151. The proteins mediate signal transduction events that play a role in the regulation of cell development, activation, growth, and motility. The 26 kDa cell surface protein is involved in adhesion, activation, proliferation, and differentiation of B cells, T cells, and other cells. On B cells CD81 is part of a complex with CD21, CD19, and Leu13. Similarly on T cells CD81 associates with CD4 and CD8 and provides a costimulatory signal with CD3. CD81 is expressed by epithelial and endothelial cells, T and B cells, as well as natural killer (NK) cells.
Additional information: Clone REA513 displays negligible binding to Fc receptors.

Alternative names

CVID6, S5.7, TAPA1, TSPAN28

Detailed product information

Technical specifications

CloneREA513
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman, non-human primate
Cross-reactivity
chimpanzee (
Pan troglodytes
)
AntigenCD81
Alternative names of antigenCVID6, S5.7, TAPA1, TSPAN28
Molecular mass of antigen [kDa]26
Distribution of antigenB cells, endothelial cells, epithelial cells, NK cells, T cells
Entrez Gene ID975
RRIDAB_2733841, AB_2751874, AB_2751844, AB_2801848, AB_2801844, AB_2857647, AB_2857617, AB_2819667, AB_2659280, AB_2659281, AB_2659282, AB_2659283, AB_2733840

Resources for CD81 Antibody, anti-human, REAfinity™

Certificates

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

References for CD81 Antibody, anti-human, REAfinity™

Publications

  1. Levy, S. (1998) CD81 (TAPA-1): a molecule involved in signal transduction and cell adhesion in the immune system. Annu. Rev. Immunol. 16: 89-109
  2. Wright, M. D. et al. (1994) The ins and outs of the transmembrane 4 superfamily. Immunol. Today 15(12): 588-594
  3. Levy, S. (1991) Structure and membrane topology of TAPA-1. J. Biol. Chem. 266(22): 14597-14602
  4. Oren, R. (1990) TAPA-1, the target of an antiproliferative antibody, defines a new family of transmembrane proteins. Mol. Cell. Biol. 10(8): 4007-4015

Related products for
CD81 Antibody, anti-human, REAfinity™

3 products available

Seems like you are coming from USA!
Do you want to visit our website in your country?