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

Name: CD314 (NKG2D) Antibody, anti-human, REAfinity™
Availability: InStock

CD314 (NKG2D) Antibody, anti-human, REAfinity™Extended ValidationRecombinant

Clone:

REA797

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

FC, MC

Alternative names:

Klrk1, KLR, NKG2D

Flow cytometry applications forCD314 (NKG2D) Antibody, anti-human, REAfinity™

APC
APC-Vio 770
Biotin
PE
PE-Vio 615
PE-Vio 770
Vio Bright B515

Conjugate:

APC

Recommended dilution:

1:50

Remark:

Cells should be stained prior to fixation, if formaldehyde is used as a fixative.

Tested:

QC tested

Products used:

130-111-999, 130-111-846

Protocols:

Cell surface flow cytometry staining protocol (PBS/EDTA/BSA 1:50)

Description:

Human peripheral blood mononuclear cells (PBMCs) were stained with CD314 (NKG2D) antibodies or with the corresponding REA Control (S) antibodies (left image) as well as with CD56 antibodies. Flow cytometry was performed using the MACSQuant^® Analyzer. The Tandem Signal Enhancer has been used to increase binding specificity of tandem-dye–conjugated antibodies. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence or 4',6-diamidino-2-phenylindole (DAPI) fluorescence, as in the case of tandem conjugates.

Mass cytometry applications forCD314 (NKG2D) Antibody, anti-human, REAfinity™

pure

Conjugate:

pure

Products used:

130-122-332

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 clones	Overlap 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.

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.

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.

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

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

Clone	REA797
Clonality	monoclonal
Isotype	recombinant human IgG1
Isotype control	REA Control Antibody (S), human IgG1
Host	human cell line
Type of antibody	Primary antibodies, Recombinant antibodies
Species	human, non-human primate
Cross-reactivity	cynomolgus monkey ( Macaca fascicularis ) , rhesus monkey ( Macaca mulatta )
Antigen	CD314 (NKG2D)
Alternative names of antigen	Klrk1, KLR, NKG2D
Molecular mass of antigen [kDa]	25
Distribution of antigen	T cells, NK cells
Entrez Gene ID	22914
RRID	AB_2657364, AB_2657365, AB_2657366, AB_2657367, AB_2657368, AB_2657369, AB_2657370, AB_2657371, AB_2657372, AB_2657373, AB_2657374, AB_2801911, AB_2922152, AB_2922117, AB_2657363

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

Certificates

Please follow this

link

to search for Certificates of Analysis (CoA) by lot number.

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

Publications

Bauer, S. et al. (1999) Activation of NK cells and T cells by NKG2D, a receptor for stress-inducible MICA. Science 285: 727-729
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
Moretta, L. and Moretta, A. (2004) Unravelling natural killer function: Triggering and inhibitory human NK receptors. EMBO J. 23: 255-259
Warren, H. S. (2005) The Eighth Human Leucocyte Differentiation Antigen (HLDA8) Workshop: Natural killer cell section report. Cell. Immunol. 236: 17-20
Maasho, K. et al. (2005)
NKG2D is a costimulatory receptor for human naive CD8
⁺
T cells.
J Immunol 174: 4480-4484
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

Applications

Extended validation