DESCRIPTION

ADAM8 is a proteolytically active member of the ADAM protease family originally described in inflammatory processes and subsequently in many systems of the body. Increased expression of ADAM8 was observed in other neoplasias, such as high-grade glioma, lung adenocarcinoma, prostate cancer, and more recently, in squamous head and neck cell carcinoma, medulloblastoma, osteosarcoma and breast cancer suggesting that ADAM8 plays an active role in tumour progression. Thus, understanding the functional role of ADAM8 in tumour biology is important.

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 (ng/mL)	SD	CV%
1	8	20.33	0.22	1.08
2	8	5.13	0.07	1.36
3	8	2.52	0.09	3.57

Inter-assay Precision

Sample	n	Mean (ng/mL)	SD	CV%
1	16	20.04	0.40	2.00
2	16	5.09	0.09	1.77
3	16	2.44	0.11	4.51

RECOVERY

The recovery of human ADAM8 spiked to three different levels throughout the range of the assay in human plasma was evaluated.

Sample Type		Average% of Expected	Range (%)
Human plasma	1:2	95	92-97
	1:4	99	90-112

LINEARITY

To assess the linearity of the assay, human plasma samples were spiked with high concentrations of human ADAM8 and diluted with the appropriate Sample Diluent to produce samples with values within the dynamic range of the assay.

		Human plasma
1:2	Average% of Expected	80
	Range (%)	73-87
1:4	Average% of Expected	104
	Range (%)	99-109
1:8	Average% of Expected	113
	Range (%)	103-123
1:16	Average% of Expected	120
	Range (%)	113-127

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human ADAM8 produced at Sunlong Medical™.