Clone:
REA366
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC
Alternative names:
GP52-40, TSPAN26

Extended validation for CD37 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 REA366
M-B371++
MB-1++
REAL335++
Cells were incubated with an excess of purified unconjugated CD37 (REA366) 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 CD37 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD37-PE (REA366, red) and counterstained with DRAQ5 (blue) as DNA stain.
View details
Fluorescence microscopy image of CD37 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD37-PE (REA366, red) and counterstained with DRAQ5 (blue) as DNA stain.
Fluorescence microscopy image of CD37 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD37-PE (REA366, red) and counterstained with DRAQ5 (blue) as DNA stain.

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 CD37. Human peripheral blood mononuclear cells (PBMCs) were stained with CD37 antibodies and with a suitable counterstaining. As a control, CD37 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 CD37. Human peripheral blood mononuclear cells (PBMCs) were stained with CD37 antibodies and with a suitable counterstaining. As a control, CD37 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 CD37. Human peripheral blood mononuclear cells (PBMCs) were stained with CD37 antibodies and with a suitable counterstaining. As a control, CD37 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 CD37. Human peripheral blood mononuclear cells (PBMCs) were stained with CD37 antibodies and with a suitable counterstaining. As a control, CD37 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 CD37. Human peripheral blood mononuclear cells (PBMCs) were stained with CD37 antibodies and with a suitable counterstaining. As a control, CD37 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 CD37 (REA366). 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 CD37 (REA366). 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 CD37 (REA366). 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 CD37 Antibody, anti-human, REAfinity™

Overview

Clone REA366 recognizes the human CD37 antigen, a multi-pass membrane protein which is also known as tetraspanin-26 (Tspan-26). CD37 is a member of the transmembrane 4 superfamily of tetraspanin proteins, which consist of 4 potential membrane-spanning regions, 2 extracellular loops, and 2 short intracytoplasmic tails. Although most tetraspanins are ubiquitous proteins, CD37 expression is nearly exclusively limited to mature B cells and B cell–derived lymphoid malignancies. B cell early progenitors, T cells, natural killer cells, and myeloid cells exhibit only minimal amounts of membrane-associated CD37. In innate immunity, CD37 interacts with pattern recognition receptor dectin-1, stabilizing dectin-1 at the macrophage cell surface, and negatively regulating proinflammatory cytokine secretion following ligand recognition. Adaptive humoral immune responses are also perturbed by CD37 ablation. In cellular immunity, CD37 negatively regulates T cell proliferation. In APCs, CD37 associates with MHC class II at the cell surface and has been shown to negatively regulate antigen presentation. CD37 has recently attracted interest as a target for monoclonal antibodies with therapeutic potential in B cell malignancies.
Additional information: Clone REA366 displays negligible binding to Fc receptors.

Alternative names

GP52-40, TSPAN26

Detailed product information

Technical specifications

CloneREA366
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hosthuman cell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
AntigenCD37
Alternative names of antigenGP52-40, TSPAN26
Molecular mass of antigen [kDa]32
Distribution of antigenB cells
Entrez Gene ID951
RRIDAB_2819463, AB_2657770, AB_2657771, AB_2657774, AB_2657775, AB_2657776, AB_2657777, AB_2657778, AB_2657779, AB_2657780, AB_2657781, AB_2657782, AB_2657783, AB_2657784, AB_2657785, AB_2928447, AB_2928446, AB_2819478

Resources for CD37 Antibody, anti-human, REAfinity™

Certificates

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References for CD37 Antibody, anti-human, REAfinity™

Publications

  1. Schwartz-Albiez, R. et al. (1988) The B cell-associated CD37 antigen (gp40-52). Structure and subcellular expression of an extensively glycosylated glycoprotein. J Immunol 140(3): 905-914
  2. Classon, B. J. et al. (1989) The primary structure of the human leukocyte antigen CD37, a species homologue of the rat MRC OX-44 antigen. J. Exp. Med. 169(4): 1497-1502
  3. Gartlan, K. H. et al. (2013) Tetraspanin CD37 contributes to the initiation of cellular immunity by promoting dendritic cell migration. Eur. J. Immunol. 43(5): 1208-1219

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