Clone:
REA631
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC, MC
Alternative names:
NKR-P1A, HNKR-P1a, CLEC5B

Extended validation for CD161 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 REA631
702228++
191B8++
DX12-
HP-3G10++
Cells were incubated with an excess of purified unconjugated CD161 (REA631) 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.

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 CD161. Human peripheral blood mononuclear cells (PBMCs) were stained with CD161 antibodies and with a suitable counterstaining. As a control, CD161 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 CD161. Human peripheral blood mononuclear cells (PBMCs) were stained with CD161 antibodies and with a suitable counterstaining. As a control, CD161 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 CD161. Human peripheral blood mononuclear cells (PBMCs) were stained with CD161 antibodies and with a suitable counterstaining. As a control, CD161 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 CD161. Human peripheral blood mononuclear cells (PBMCs) were stained with CD161 antibodies and with a suitable counterstaining. As a control, CD161 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 CD161. Human peripheral blood mononuclear cells (PBMCs) were stained with CD161 antibodies and with a suitable counterstaining. As a control, CD161 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 CD161 (REA631). 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 CD161 (REA631). 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 CD161 (REA631). 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 CD161 Antibody, anti-human, REAfinity™

Overview

Clone REA631 recognizes the human CD161 antigen, a 80 kDa single-pass type II membrane protein, which is also known as natural killer cell surface protein P1A (NKR-P1A) or C-type lectin domain family 5 member B (CLEC5B). CD161 is expressed on most natural killer (NK) cells and subsets of T cells. In peripheral blood, CD161 is preferentially expressed on T cells of memory phenotype, but it can also be found on subsets of thymocytes and fetal liver T cells. CD161 has been implicated in triggering NK-mediated cytotoxicity, contributing to target cell recognition by NK cells.
Additional information: Clone REA631 displays negligible binding to Fc receptors.

Alternative names

NKR-P1A, HNKR-P1a, CLEC5B

Detailed product information

Technical specifications

CloneREA631
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
AntigenCD161
Alternative names of antigenNKR-P1A, HNKR-P1a, CLEC5B
Molecular mass of antigen [kDa]25
Distribution of antigenT cells, NK cells, thymocytes, liver
Entrez Gene ID3820
RRIDAB_2751137, AB_2751202, AB_2751136, AB_2751201, AB_2751135, AB_2726467, AB_2726190, AB_2751817, AB_2751761, AB_2811391, AB_2811388, AB_2811536, AB_2811527, AB_2801923, AB_2751203

References for CD161 Antibody, anti-human, REAfinity™

Publications

  1. Lanier, L. L. et al. (1994) Human NKR-P1A. A disulfide-linked homodimer of the C-type lectin superfamily expressed by a subset of NK and T lymphocytes. J. Immunol. 153: 2417-2428
  2. Pozo, D. et al. (2006) CD161 (human NKRP1A) signaling in NK cells involves the activation of acid sphingomyelinase. J. Immunol. 176: 2397-2406
  3. Takahashi, T. et al. (2006) Expression of CD161 (NKR-P1A) defines subsets of human CD4 and CD8 T cells with different functional activities.​ J. Immunol. 176: 211-216

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