Clone:
REA168
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
MC, FC
Alternative names:
KIR3DL1, KIR3DS1, HLA-BW4-specific inhibitory NK cell receptor, MHC class I NK cell receptor, Natural killer-associated transcript 3 (NKAT-3, NKAT3), p70 natural killer cell receptor clones CL-2/CL-11 (p70 NK receptor CL-2/CL-11), KIRp70/NKB1

Extended validation for CD158e1/e2 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 REA168
Z27.3.7++
Cells were incubated with an excess of purified unconjugated CD158e1/e2 (REA168) 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 CD158e1/e2. Human peripheral blood mononuclear cells (PBMCs) were stained with CD158e1/e2 antibodies and with a suitable counterstaining. As a control, CD158e1/e2 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 CD158e1/e2. Human peripheral blood mononuclear cells (PBMCs) were stained with CD158e1/e2 antibodies and with a suitable counterstaining. As a control, CD158e1/e2 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 CD158e1/e2. Human peripheral blood mononuclear cells (PBMCs) were stained with CD158e1/e2 antibodies and with a suitable counterstaining. As a control, CD158e1/e2 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 CD158e1/e2. Human peripheral blood mononuclear cells (PBMCs) were stained with CD158e1/e2 antibodies and with a suitable counterstaining. As a control, CD158e1/e2 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 CD158e1/e2 (REA168). 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 CD158e1/e2 (REA168). 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 CD158e1/e2 (REA168). 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 CD158e1/e2 Antibody, anti-human, REAfinity™

Overview

Clone REA168 recognizes the CD158e1/e2 antigens, which are single-pass type I membrane proteins. CD158e is expressed on CD56
dim
CD16
+
natural killer (NK) cells and CD8
+
T cells. Killer cell immunoglobulin-like receptors (KIRs) contribute to the regulation of NK cell–mediated cytotoxicity. The KIR proteins are classified by the number of extracellular immunoglobulin domains (2D or 3D) and by whether they have a long or short cytoplasmic domain. According to the length of their cytoplasmic tail, KIRs can be further subdivided into inhibitory KIRs and activating KIRs. The CD158e1 antigen is also known as KIR3DL1 (KIR, three Ig like domain extracellular domains, long cytoplasmic tail, 1) and the CD158e2 antigen as KIR3DS1 (KIR, three extracellular Ig like domains, short cytoplasmic tail, 1). Within the KIR3DL1/S1 gene locus, CD158e2 represents a conserved allelic variant and displays unique features in comparison to the highly polymorphic CD158e1 allele. CD158e1 provides an inhibitory signal of NK cell lytic activity upon interaction with its specific ligand, HLA-Bw4. Despite the lack of formal evidence for an interaction of CD158e2 with HLA-Bw4 or any other HLA class I subtype, a growing number of associations between the presence of CD158e2 and the outcome of viral infections have been described. Especially, the potential protective role of CD158e2 in combination with HLA-Bw4-I80 in the context of HIV-1 infection has been studied intensively.
Additional information: Clone REA168 displays negligible binding to Fc receptors.

Alternative names

KIR3DL1, KIR3DS1, HLA-BW4-specific inhibitory NK cell receptor, MHC class I NK cell receptor, Natural killer-associated transcript 3 (NKAT-3, NKAT3), p70 natural killer cell receptor clones CL-2/CL-11 (p70 NK receptor CL-2/CL-11), KIRp70/NKB1

Detailed product information

Technical specifications

CloneREA168
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
AntigenCD158e1/e2
Alternative names of antigenKIR3DL1, KIR3DS1, HLA-BW4-specific inhibitory NK cell receptor, MHC class I NK cell receptor, Natural killer-associated transcript 3 (NKAT-3, NKAT3), p70 natural killer cell receptor clones CL-2/CL-11 (p70 NK receptor CL-2/CL-11), KIRp70/NKB1
Molecular mass of antigen [kDa]44(average of subunits)
Distribution of antigenNK cells, T cells
Entrez Gene ID3811, 3813
RRIDAB_2857825, AB_2857818, AB_2857668, AB_2857645, AB_2655345, AB_2655346, AB_2655349, AB_2655350, AB_2655351, AB_2655352, AB_2655353, AB_2655354, AB_2655355, AB_2655356, AB_2655359, AB_2655360, AB_2801865

Resources for CD158e1/e2 Antibody, anti-human, REAfinity™

Documents and Protocols

App notes/customer reports

Cross-reactivity of KIR antibodies

References for CD158e1/e2 Antibody, anti-human, REAfinity™

Publications

  1. Carr, W. H. et al. (2005) KIR3DL1 polymorphisms that affect NK cell inhibition by HLA-Bw4 ligand. J. Immunol. 175(8): 5222-5229
  2. Moretta, A. et al. (2001) Activating receptors and coreceptors involved in human natural killer cell-mediated cytolysis. Annu. Rev. Immunol. 19: 197-223
  3. Körner, C. et al. (2012) Role of KIR3DS1 in human diseases. Front Immunol 3: 326

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