DESCRIPTION

NCOA6 (Nuclear receptor coactivator 6), also named as AIB3 or ANTP, is a 2063 amino acid protein, which contains two LXXLL motifs. NCOA6 localizes in the nucleus and is highly expressed in brain, prostate, testis and ovary. NCOA6 directly binds nuclear receptors and stimulates the transcriptional activities in a hormone-dependent fashion.

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	2,806.3	42.7	1.5
2	8	712.5	13.5	1.9
3	8	367.1	10.3	2.8

Inter-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	16	2,795.7	51.0	1.8
2	16	699.5	18.9	2.7
3	16	364.2	12.2	3.3

RECOVERY

The recovery of human NCOA6 spiked to three different levels throughout the range of the assay in cell lysate was evaluated.

Sample Type		Average% of Expected	Range (%)
Cell lysate	1:8	122	120-125
	1:16	121	118-128

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.

		Cell lysate
1:2	Average% of Expected	100
	Range (%)	-
1:4	Average% of Expected	97
	Range (%)	96-99
1:8	Average% of Expected	88
	Range (%)	85-92
1:16	Average% of Expected	74
	Range (%)	70-77

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human Human NCOA6 One-Step ELISA Kit produced at Sunlong Medical™.