DESCRIPTION

NKp46, also known as NCR1 or CD335, is an Ig-like superfamily cell surface receptor that is highly conserved in mammals. NKp46 is a type I transmembrane glycoprotein consisting of two extracellular Ig-like domains, a transmembrane domain, and an intracellular tail. It is expressed on NK cells, rare T-cell subsets, and a mucosal population of NKp46+ innate lymphoid cells. NKp46 is the major triggering receptor involved in natural cytotoxicity.

PRINCIPLE OF THE ASSAY

An anti-tag antibody is pre-coated onto the bottom of wells. After adding antigen or samples, Tagged-Capture antibody and HRP-conjugated detection antibody, a sandwich complex is formed in the solution. TMB acts as a HRP substrate, and the solution color will change from colorless to blue. A stop solution containing sulfuric acid turns the solution yellow. The color intensity is proportional to the quantity of bound protein, which is measurable at 450 nm with the correction wavelength set at 630 nm.

PRECISION

Intra-assay Precision (Precision within an assay) Three samples of known concentration were tested 8 times on one plate to assess intra-assay precision. Inter-assay Precision (Precision between assays) Three samples of known concentration were tested in 16 separate assays to assess inter-assay precision.

Intra-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	8	1,042.6	6.1	0.6
2	8	239.0	3.6	1.5
3	8	112.3	2.9	2.6

Inter-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	16	1,045.9	10.1	1.0
2	16	234.3	6.6	2.8
3	16	111.2	2.9	2.6

RECOVERY

The recovery of human NCR1 spiked to three different levels throughout the range of the assay in various matrices was evaluated.

Sample Type		Average% of Expected	Range (%)
Human plasma	1:2	95	88-99
	1:4	101	98-105
Cell culture supernatant	1:4	101	85-112
	1:8	106	93-114

LINEARITY

To assess the linearity of the assay, samples were diluted with the appropriate Sample Diluent to produce samples with values within the dynamic range of the assay.

		Human plasma	Cell culture supernatant
1:2	Average% of Expected	100	100
	Range (%)	-	-
1:4	Average% of Expected	111	111
	Range (%)	99-123	108-113
1:8	Average% of Expected	108	111
	Range (%)	92-125	106-116
1:16	Average% of Expected	-	120
	Range (%)	-	117-122

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human NCR1 produced at Sunlong Medical™.