Clone:
REA340
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC
Alternative names:
SIGLEC-2, B-lymphocyte cell adhesion molecule (BL-CAM), Sialic acid-binding Ig-like lectin 2 (Siglec-2), T cell surface antigen Leu-14

Extended validation for CD22 Antibody, anti-human, REAfinity™

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
View details
Fluorescence microscopy image of CD22 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD22-PE (REA340, red) and counterstained with DRAQ5 (blue) as DNA stain.
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Fluorescence microscopy image of CD22 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD22-PE (REA340, red) and counterstained with DRAQ5 (blue) as DNA stain.
Fluorescence microscopy image of CD22 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD22-PE (REA340, red) and counterstained with DRAQ5 (blue) as DNA stain.
View details
Overlay histogram showing flow cytometric analysis of human CD22 knockout cells. Wild type (red) and knockout cells (blue) were stained with CD22-PE (REA340). Flow cytometry was performed with the MACSQuant
®
Analyzer. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.
Overlay histogram showing flow cytometric analysis of human CD22 knockout cells. Wild type (red) and knockout cells (blue) were stained with CD22-PE (REA340). Flow cytometry was performed with the MACSQuant
®
Analyzer. Cell debris and dead cells 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 CD22. Human peripheral blood mononuclear cells (PBMCs) were stained with CD22 antibodies and with a suitable counterstaining. As a control, CD22 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 CD22. Human peripheral blood mononuclear cells (PBMCs) were stained with CD22 antibodies and with a suitable counterstaining. As a control, CD22 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 CD22. Human peripheral blood mononuclear cells (PBMCs) were stained with CD22 antibodies and with a suitable counterstaining. As a control, CD22 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 CD22. Human peripheral blood mononuclear cells (PBMCs) were stained with CD22 antibodies and with a suitable counterstaining. As a control, CD22 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 CD22. Human peripheral blood mononuclear cells (PBMCs) were stained with CD22 antibodies and with a suitable counterstaining. As a control, CD22 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 CD22. Human peripheral blood mononuclear cells (PBMCs) were stained with CD22 antibodies and with a suitable counterstaining. As a control, CD22 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 CD22. Human peripheral blood mononuclear cells (PBMCs) were stained with CD22 antibodies and with a suitable counterstaining. As a control, CD22 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 CD22. Human peripheral blood mononuclear cells (PBMCs) were stained with CD22 antibodies and with a suitable counterstaining. As a control, CD22 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 CD22. Human peripheral blood mononuclear cells (PBMCs) were stained with CD22 antibodies and with a suitable counterstaining. As a control, CD22 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 CD22 (REA340). 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 CD22 (REA340). 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 CD22 (REA340). 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 CD22 (REA340). 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 CD22 (REA340). 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 CD22 Antibody, anti-human, REAfinity™

Overview

Clone REA340 recognizes the human CD22 antigen, a single-pass type I membrane protein which is also known as sialic acid-binding Ig-like lectin 2 (Siglec-2). In peripheral blood, cell surface expression of CD22 is found on mature B cells, but is lost during terminal differentiation to plasma cells. In bone marrow, surface expression of CD22 can be detected from the pre–B cell stage on. Cytoplasmic expression of CD22 can also be found in late pro–B cells and early–B cells. CD22 plays a strong role in cell interactions and B cell activation. It is a regulatory molecule that prevents the overactivation of the immune system and the development of autoimmune diseases. CD22 has emerged as an ideal target for monoclonal antibody based therapy of B cell malignancies including most lymphomas and many leukemias.
Additional information: Clone REA340 displays negligible binding to Fc receptors.

Alternative names

SIGLEC-2, B-lymphocyte cell adhesion molecule (BL-CAM), Sialic acid-binding Ig-like lectin 2 (Siglec-2), T cell surface antigen Leu-14

Detailed product information

Technical specifications

CloneREA340
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hosthuman cell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
AntigenCD22
Alternative names of antigenSIGLEC-2, B-lymphocyte cell adhesion molecule (BL-CAM), Sialic acid-binding Ig-like lectin 2 (Siglec-2), T cell surface antigen Leu-14
Molecular mass of antigen [kDa]93
Distribution of antigenB cells, bone marrow
Entrez Gene ID933
RRIDAB_2752186, AB_2802028, AB_2802011, AB_2819604, AB_2819589, AB_2904853, AB_2904852, AB_2656361, AB_2656362, AB_2656363, AB_2656364, AB_2656365, AB_2656366, AB_2656369, AB_2656370, AB_2904851, AB_2904850, AB_2904849, AB_2904848, AB_2904855, AB_2904854, AB_2752208

Resources for CD22 Antibody, anti-human, REAfinity™

Certificates

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

References for CD22 Antibody, anti-human, REAfinity™

Publications

  1. Stamenkovic, I. et al. (1990) The B-cell antigen CD22 mediates monocyte and erythrocyte adhesion. Nature 345(6270): 74-77
  2. Wilson, G. L. et al. (1991) cDNA cloning of the B cell membrane protein CD22: a mediator of B-B cell interactions. J. Exp. Med. 173(1): 137-146
  3. Sullivan-Chang, L. et al. (2013) Targeting CD22 in B-cell malignancies: current status and clinical outlook. BioDrugs 27(4): 293-304

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