CD337 (NKp30) Antibody, anti-human, REAfinity™

Name: CD337 (NKp30) Antibody, anti-human, REAfinity™
Availability: InStock

CD337 (NKp30) Antibody, anti-human, REAfinity™Extended ValidationRecombinant

Clone:

REA823

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

FC

Alternative names:

NCR3, 1C7, LY117, MALS, NKp30

Flow cytometry applications forCD337 (NKp30) Antibody, anti-human, REAfinity™

APC
Biotin
PE
PE-Vio 615
PE-Vio 770
Vio Bright B515
Vio Bright V423

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:

130-112-503, 130-112-431

Protocols:

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

Description:

Human peripheral blood mononuclear cells (PBMCs) were stained with CD337 (NKp30) 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.

Extended validation for CD337 (NKp30) 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 REA823
AF29-4D12	++
P30-15	++
Cells were incubated with an excess of purified unconjugated CD337 (NKp30) (REA823) 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 CD337 (NKp30). Human peripheral blood mononuclear cells (PBMCs) were stained with CD337 (NKp30) antibodies and with a suitable counterstaining. As a control, CD337 (NKp30) 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 CD337 (NKp30). Human peripheral blood mononuclear cells (PBMCs) were stained with CD337 (NKp30) antibodies and with a suitable counterstaining. As a control, CD337 (NKp30) 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 CD337 (NKp30). Human peripheral blood mononuclear cells (PBMCs) were stained with CD337 (NKp30) antibodies and with a suitable counterstaining. As a control, CD337 (NKp30) 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 CD337 (NKp30). Human peripheral blood mononuclear cells (PBMCs) were stained with CD337 (NKp30) antibodies and with a suitable counterstaining. As a control, CD337 (NKp30) 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 CD337 (NKp30). Human peripheral blood mononuclear cells (PBMCs) were stained with CD337 (NKp30) antibodies and with a suitable counterstaining. As a control, CD337 (NKp30) 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 CD337 (NKp30) (REA823). 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 CD337 (NKp30) (REA823). 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 CD337 (NKp30) Antibody, anti-human, REAfinity™

Overview

Clone REA823 recognizes the CD337 (NKp30) receptor, which is a 30 kDa type I membrane glycoprotein characterized by a single V-type Ig-like domain in the extracellular portion. CD337 is found almost exclusively on the surface of resting or activated natural killer (NK) cells and is a member of the natural cytotoxicity receptor (NCR) family which trigger cytotoxicity in NK cells. CD337 is directly involved in target cell recognition and lysis and is expressed by all resting or activated NK cells and by a small subset of T cells. Its expression density parallels that of CD336 (NKp44). It seems to be involved in killing of cells which are lysed independently from CD336 (NKp44) and CD335 (NKp46).

Additional information: Clone REA823 displays negligible binding to Fc receptors.

Alternative names

NCR3, 1C7, LY117, MALS, NKp30

Detailed product information

Technical specifications

Clone	REA823
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
Cross-reactivity	cynomolgus monkey ( Macaca fascicularis ) , rhesus monkey ( Macaca mulatta )
Antigen	CD337 (NKp30)
Alternative names of antigen	NCR3, 1C7, LY117, MALS, NKp30
Molecular mass of antigen [kDa]	20
Distribution of antigen	NK cells
Entrez Gene ID	259197
RRID	AB_2657619, AB_2657620, AB_2657621, AB_2657622, AB_2657623, AB_2657624, AB_2657625, AB_2928426, AB_2928425, AB_2657626, AB_2657627, AB_2921829, AB_2921848, AB_2657618

Resources for CD337 (NKp30) Antibody, anti-human, REAfinity™

Certificates

Please follow this

link

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

References for CD337 (NKp30) Antibody, anti-human, REAfinity™

Publications

Pende, D. et al. (1999) Identification and molecular characterization of Nkp30, a novel triggering receptor involved in natural cytotoxicity mediated by human natural killer cells. J. Exp. Med. 190: 1505-1516
Sivori, S. et al. (2000) 2B4 functions as a co-receptor in human NK cell activation. Eur. J. Immunol. 30: 787-793
Moretta, A. et al. (2001) Activating receptors and coreceptors involved in human natural killer cell-mediated cytolysis. Annu. Rev. Immunol. 19: 197-223
Moretta, L. and Moretta, A. (2004) Unravelling natural killer function: Triggering and inhibitory human NK receptors. EMBO J. 23: 255-259

Applications

Extended validation