Clone:
REA196
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MC
Alternative names:
NCAM1, MSK39, NCAM

Extended validation for CD56 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 REA196
NCAM16.2++
TULY56++
5.1H11++
AF12-7H3-
C5.9++
N901/NKH1++
CMSSB+
B159-
HCD56-
MEM188-
Cells were incubated with an excess of purified unconjugated CD56 (REA196) 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 CD56 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD56-PE (REA196, red) and counterstained with DRAQ5 (blue) as DNA stain.
View details
Fluorescence microscopy image of CD56 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD56-PE (REA196, red) and counterstained with DRAQ5 (blue) as DNA stain.
Fluorescence microscopy image of CD56 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD56-PE (REA196, red) and counterstained with DRAQ5 (blue) as DNA stain.
View details
Overlay histogram showing flow cytometric analysis of CD56 knockout cells. Wild type (red) and knockout cells (blue) were stained with CD56-PE, clone (REA196). 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 CD56 knockout cells. Wild type (red) and knockout cells (blue) were stained with CD56-PE, clone (REA196). 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 CD56. Human peripheral blood mononuclear cells (PBMCs) were stained with CD56 antibodies and with a suitable counterstaining. As a control, CD56 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 CD56. Human peripheral blood mononuclear cells (PBMCs) were stained with CD56 antibodies and with a suitable counterstaining. As a control, CD56 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 CD56. Human peripheral blood mononuclear cells (PBMCs) were stained with CD56 antibodies and with a suitable counterstaining. As a control, CD56 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 CD56. Human peripheral blood mononuclear cells (PBMCs) were stained with CD56 antibodies and with a suitable counterstaining. As a control, CD56 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 CD56. Human peripheral blood mononuclear cells (PBMCs) were stained with CD56 antibodies and with a suitable counterstaining. As a control, CD56 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 CD56. Human peripheral blood mononuclear cells (PBMCs) were stained with CD56 antibodies and with a suitable counterstaining. As a control, CD56 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 CD56. Human peripheral blood mononuclear cells (PBMCs) were stained with CD56 antibodies and with a suitable counterstaining. As a control, CD56 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 CD56. Human peripheral blood mononuclear cells (PBMCs) were stained with CD56 antibodies and with a suitable counterstaining. As a control, CD56 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 CD56. Human peripheral blood mononuclear cells (PBMCs) were stained with CD56 antibodies and with a suitable counterstaining. As a control, CD56 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 CD56. Human peripheral blood mononuclear cells (PBMCs) were stained with CD56 antibodies and with a suitable counterstaining. As a control, CD56 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 CD56. Human peripheral blood mononuclear cells (PBMCs) were stained with CD56 antibodies and with a suitable counterstaining. As a control, CD56 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 CD56. Human peripheral blood mononuclear cells (PBMCs) were stained with CD56 antibodies and with a suitable counterstaining. As a control, CD56 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 CD56 (REA196). 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 CD56 (REA196). 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 CD56 (REA196). 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 CD56 Antibody, anti-human, REAfinity™

Overview

Clone REA196 recognizes the human CD56 antigen, a glycoprotein of the Ig-superfamily, also known as neural cell adhesion molecule (NCAM), which is expressed in blood on practically all resting and activated NK cells and on a minor subset of CD3
+
T cells. CD56 is also expressed in brain (cerebellum and cortex) and at neuromuscular junctions. Certain large granular lymphocyte (LGL) leukemias, small-cell lung carcinomas, neuronal-derived tumors, myelomas, and myeloid leukemias also express CD56.
Additional information: Clone REA196 displays negligible binding to Fc receptors.

Alternative names

NCAM1, MSK39, NCAM

Detailed product information

Technical specifications

CloneREA196
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
)
,
rhesus monkey (
Macaca mulatta
)
,
cynomolgus monkey (
Macaca fascicularis
)
,
olive baboon (
Papio anubis
)
AntigenCD56
Alternative names of antigenNCAM1, MSK39, NCAM
Molecular mass of antigen [kDa]92
Distribution of antigenNK cells, T cells
Entrez Gene ID4684
RRIDAB_2726695, AB_2726367, AB_2726090, AB_2726365, AB_2726088, AB_2726780, AB_2726696, AB_2726368, AB_2726091, AB_2733135, AB_2733136, AB_2733037, AB_2733038, AB_2726781, AB_2726697, AB_2726366, AB_2726089, AB_2819377, AB_2819374, AB_2658728, AB_2726779

References for CD56 Antibody, anti-human, REAfinity™

Publications

  1. Camp, B. V. et al. (1990) Plasma cells in multiple myeloma express a natural killer cell- associated antigen: CD56 (NKH-1; Leu-19). Blood 76: 377-382
  2. Vivier, E. et al. (2018) Anti-NKG2A mAb Is a Checkpoint Inhibitor that Promotes Anti-tumor Immunity by Unleashing Both T and NK Cells. Cell 175(7): 1731-1743
  3. Boin, F. et al. (2017) Flow cytometric discrimination of seven lineage markers by using two fluorochromes. PLoS One 12(11)
  4. Petty, R. D. et al. (2016) MicroRNAs hsa-miR-99b, hsa-miR-330, hsa-miR-126 and hsa-miR-30c: potential diagnostic biomarkers in natural killer (NK) cells of patients with chronic fatigue syndrome (CFS)/ myalgic encephalomyelitis (ME). PLoS One 11: e0150904
  5. Dong, P. et al. (2017) Simultaneous detection of decidual Th1/Th2 and NK1/NK2 immunophenotyping in unknown recurrent miscarriage using 8-color flow cytometry with FSC/Vt extended strategy. Biosci. Rep. 37(3)
  6. Canté-Barrett, K. et al. (2017)
    Loss of CD44
    dim
    expression from early progenitor cells marks T-cell lineage commitment in the human thymus.
    Front Immunol 8: 32
  7. Croci, S. et al. (2018) Higher frequencies of lymphocytes expressing the natural killer group 2D receptor in patients with Behçet disease. Front Immunol 9: 2157
  8. Delso-Vallejo, M. et al. (2017)
    Influence of irradiated peripheral blood mononuclear cells on both
    ex vivo
    proliferation of human natural killer cells and change in cellular property.
    Front Immunol 8
  9. Grant, M. D. et al. (2018) Natural Killer Cells Adapt to Cytomegalovirus Along a Functionally Static Phenotypic Spectrum in Human Immunodeficiency Virus Infection. Front Immunol 9: 2494
  10. Popkin, D. L. et al. (2018)
    In vitro
    evidence that combination therapy with CD16-bearing NK-92 cells and FDA-approved Alefacept can selectively target the latent HIV reservoir in CD4
    +
    CD2
    hi
    memory T cells.
    Front Immunol 9: 2552
  11. Veluchamy, J. et al. (2017)
    In vivo
    efficacy of umbilical cord blood stem cell-derived NK cells in the treatment of metastatic colorectal cancer.
    Front Immunol 8: 87

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