CD335 (NKp46) Antibody, anti-human, REAfinity™

Name: CD335 (NKp46) Antibody, anti-human, REAfinity™
Availability: InStock

CD335 (NKp46) Antibody, anti-human, REAfinity™Extended ValidationRecombinant

Clone:

REA808

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

FC, MICS, IF, IHC, MC

Alternative names:

NK-p46, hNKp46, NCR1, LY94

Flow cytometry applications forCD335 (NKp46) Antibody, anti-human, REAfinity™

APC
Biotin
PE
PE-Vio 615
PE-Vio 770
Vio Bright B515
Vio Bright R720

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:

130-112-280, 130-112-122

Protocols:

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

Description:

Human peripheral blood mononuclear cells (PBMCs) were stained with CD335 (NKp46) antibodies or with the corresponding REA Control (S) antibodies (left images) 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.

MICS (MACSima Imaging Cyclic Staining) applications forCD335 (NKp46) Antibody, anti-human, REAfinity™

APC
PE

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

PFA

Tested:

during development

Species tested:

human

Products used:

130-112-280, 130-112-122

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Mass cytometry applications forCD335 (NKp46) Antibody, anti-human, REAfinity™

pure

Conjugate:

pure

Products used:

130-124-522

Extended validation for CD335 (NKp46) 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 REA808
9E2	++
195314	++
Cells were incubated with an excess of purified unconjugated CD335 (NKp46) (REA808) 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 CD335 (NKp46). Human peripheral blood mononuclear cells (PBMCs) were stained with CD335 (NKp46) antibodies and with a suitable counterstaining. As a control, CD335 (NKp46) 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 CD335 (NKp46). Human peripheral blood mononuclear cells (PBMCs) were stained with CD335 (NKp46) antibodies and with a suitable counterstaining. As a control, CD335 (NKp46) 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 CD335 (NKp46). Human peripheral blood mononuclear cells (PBMCs) were stained with CD335 (NKp46) antibodies and with a suitable counterstaining. As a control, CD335 (NKp46) 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 CD335 (NKp46). Human peripheral blood mononuclear cells (PBMCs) were stained with CD335 (NKp46) antibodies and with a suitable counterstaining. As a control, CD335 (NKp46) 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 CD335 (NKp46). Human peripheral blood mononuclear cells (PBMCs) were stained with CD335 (NKp46) antibodies and with a suitable counterstaining. As a control, CD335 (NKp46) 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 CD335 (NKp46). Human peripheral blood mononuclear cells (PBMCs) were stained with CD335 (NKp46) antibodies and with a suitable counterstaining. As a control, CD335 (NKp46) 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 CD335 (NKp46) (REA808). 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 CD335 (NKp46) (REA808). 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 CD335 (NKp46) Antibody, anti-human, REAfinity™

Overview

Clone REA808 recognizes the CD335 antigen, a type I membrane glycoprotein also known as NKp46. CD335 is a member of the natural cytotoxicity receptor (NCR) family which trigger cytotoxicity in natural killer (NK) cells and mediates cell activation. CD335 is directly involved in target cell recognition and lysis and is exclusively expressed on CD3

^–

CD56

⁺

NK cells, suggesting it to be a universal marker for NK cells.

Additional information: Clone REA808 displays negligible binding to Fc receptors.

Alternative names

NK-p46, hNKp46, NCR1, LY94

Detailed product information

Technical specifications

Clone	REA808
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
Antigen	CD335 (NKp46)
Alternative names of antigen	NK-p46, hNKp46, NCR1, LY94
Molecular mass of antigen [kDa]	32
Distribution of antigen	NK cells
Entrez Gene ID	9437
RRID	AB_2657590, AB_2657591, AB_2657592, AB_2657593, AB_2657594, AB_2657595, AB_2657596, AB_2657597, AB_2734017, AB_2734018, AB_2657598, AB_2657599, AB_2811677, AB_2904958, AB_2904957

Resources for CD335 (NKp46) Antibody, anti-human, REAfinity™

Certificates

Please follow this

link

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

References for CD335 (NKp46) Antibody, anti-human, REAfinity™

Publications

Pessino, A. et al. (1998) Molecular cloning of NKp46: A novel member of the immunoglobulin superfamily involved in triggering of natural cytotoxicity. J. Exp. Med. 188: 953-960
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
Lai, C. B. et al. (2012) Role of runt-related transcription factor 3 (RUNX3) in transcription regulation of natural cytotoxicity receptor 1 (NCR1/NKp46), an activating natural killer (NK) cell receptor. J. Biol. Chem. 287(10): 7324-7334

Applications

Extended validation