Clone:
REA797
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MC
Alternative names:
Klrk1, KLR, NKG2D

Extended validation for CD314 (NKG2D) 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 REA797
149810++
1D11++
5C6++
BAT221++
Cells were incubated with an excess of purified unconjugated CD314 (NKG2D) (REA797) 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 CD304 (BDCA-4). Human peripheral blood mononuclear cells (PBMCs) were stained with CD304 (BDCA-4) antibodies and with a suitable counterstaining. As a control, CD304 (BDCA-4) 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 CD304 (BDCA-4). Human peripheral blood mononuclear cells (PBMCs) were stained with CD304 (BDCA-4) antibodies and with a suitable counterstaining. As a control, CD304 (BDCA-4) 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 CD304 (BDCA-4). Human peripheral blood mononuclear cells (PBMCs) were stained with CD304 (BDCA-4) antibodies and with a suitable counterstaining. As a control, CD304 (BDCA-4) 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 CD304 (BDCA-4). Human peripheral blood mononuclear cells (PBMCs) were stained with CD304 (BDCA-4) antibodies and with a suitable counterstaining. As a control, CD304 (BDCA-4) 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 CD304 (BDCA-4). Human peripheral blood mononuclear cells (PBMCs) were stained with CD304 (BDCA-4) antibodies and with a suitable counterstaining. As a control, CD304 (BDCA-4) 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 CD304 (BDCA-4). Human peripheral blood mononuclear cells (PBMCs) were stained with CD304 (BDCA-4) antibodies and with a suitable counterstaining. As a control, CD304 (BDCA-4) 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 CD314 (NKG2D) (REA797). 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 CD314 (NKG2D) (REA797). 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 CD314 (NKG2D) (REA797). 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 CD314 (NKG2D) Antibody, anti-human, REAfinity™

Overview

Clone REA797 recognizes CD314 (NKG2D), a cell surface receptor of the type II transmembrane receptor NKG2 family. It is expressed on NK cells, CD8
+
α/β T cells, and γ/δ T cells. Though NKG2D itself possesses no intrinsic signaling capability, ligand engagement while associated to the adaptor proteins DAP10 or DAP12 induces cytolytic activation in NK cells as well as costimulation in certain T cell populations. Ligands for NKG2D are stress-inducible MHC class I–related MICA, MICB, and UL16–binding proteins, expressed predominantly on epithelial cells and numerous carcinomas.
Additional information: Clone REA797 displays negligible binding to Fc receptors.

Alternative names

Klrk1, KLR, NKG2D

Detailed product information

Technical specifications

CloneREA797
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
cynomolgus monkey (
Macaca fascicularis
)
,
rhesus monkey (
Macaca mulatta
)
AntigenCD314 (NKG2D)
Alternative names of antigenKlrk1, KLR, NKG2D
Molecular mass of antigen [kDa]25
Distribution of antigenT cells, NK cells
Entrez Gene ID22914
RRIDAB_2657364, AB_2657365, AB_2657366, AB_2657367, AB_2657368, AB_2657369, AB_2657370, AB_2657371, AB_2657372, AB_2657373, AB_2657374, AB_2801911, AB_2657363

References for CD314 (NKG2D) Antibody, anti-human, REAfinity™

Publications

  1. Bauer, S. et al. (1999) Activation of NK cells and T cells by NKG2D, a receptor for stress-inducible MICA. Science 285: 727-729
  2. Groh, V. et al. (2003) Stimulation of T cell autoreactivity by anomalous expression of NKG2D and its MIC ligands in rheumatoid arthritis. Proc. Natl. Acad. Sci. U.S.A. 100(16): 9452-9457
  3. Moretta, L. and Moretta, A. (2004) Unravelling natural killer function: Triggering and inhibitory human NK receptors. EMBO J. 23: 255-259
  4. Warren, H. S. (2005) The Eighth Human Leucocyte Differentiation Antigen (HLDA8) Workshop: Natural killer cell section report. Cell. Immunol. 236: 17-20
  5. Maasho, K. et al. (2005)
    NKG2D is a costimulatory receptor for human naive CD8
    +
    T cells.
    J. Immunol. 174: 4480-4484
  6. Chaput, N. et al. (2013)
    Phase I clinical trial combining imatinib mesylate and IL-2: HLA-DR
    +
    NK cell levels correlate with disease outcome.
    Oncoimmunology 2(2): e23080

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