DESCRIPTION

SERPINE1, also named as Plasminogen activator inhibitor type 1 (PAI-1), is a member of the serine protease inhibitor (SERPIN) superfamily. It is produced by the vascular endothelium, the liver, the monocytes/macrophagues, the platelets and the adipose tissue. High plasma levels of PAI-1 have been associated with an increased risk of suffering cardiovascular diseases. It is implicated in the pathogenesis of obesity, insulin resistance and type 2 diabetes. In several tumor types, SERPINE1 expression is up-regulated and it has been described as a poor prognostic marker. Besides its prognostic value, SERPINE1 expression has been validated as a marker for therapy decision making in patients with node-negative breast cancer. Defects in this gene are the cause of plasminogen activator inhibitor-1 deficiency (PAI-1 deficiency), and high concentrations of the gene product are associated with thrombophilia.

PRINCIPLE OF THE ASSAY

Sandwich ELISA structure (Detection antibody labeled with HRP) A capture antibody is pre-coated onto the bottom of wells which binds to analyte of interest. A detection antibody labeled with HRP also binds to the analyte. TMB acts as the HRP substrate and the solution color will change from colorless to blue. A stop solution containing sulfuric acid turns 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 20 times on one plate to assess intra-assay precision. Inter-assay Precision (Precision between assays) Three samples of known concentration were tested in 24 separate assays to assess inter-assay precision.

Intra-assay Precision

Sample	n	Mean (ng/mL)	SD	CV%
1	20	1.0	0.05	4.8
2	20	4.8	0.06	1.3
3	20	17.6	0.43	2.4

Inter-assay Precision

Sample	n	Mean (ng/mL)	SD	CV%
1	24	1.4	0.04	2.6
2	24	5.2	0.13	2.5
3	24	18.4	0.36	1.9

RECOVERY

The recovery of human SERPINE1 spiked to three different levels throughout the range of the assay in human samples were evaluated.

Sample Type		Average% of Expected	Range (%)
Human plasma	1:300	84	72-98
	1:600	85	74-99
Human serum	1:160	87	79-98
	1:320	85	77-98
Cell culture supernatant	1:30	91	84-98
	1:60	89	88-91
Cell lysate	1:16	87	80-95
	1:32	89	87-93

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. (The plasma and serum samples were initially diluted 1:10, cell culture supernatant was initially diluted 1:2）

		Human plasma(Sample Diluent PT4)	Human serum(Sample Diluent PT4)	Cell culturesupernatant (SampleDiluent PT 1)	Cell lysate (SampleDiluent PT 1)
1:2	Average% of Expected	95	100	95	84
	Range (%)	93-98	95-103	94-96	82-85
1:4	Average% of Expected	104	100	98	99
	Range (%)	104-105	100-101	96-100	92-106
1:8	Average% of Expected	100	100	101	100
	Range (%)	99-101	100-101	100-103	100-101
1:16	Average% of Expected	98	99	105	106
	Range (%)	98-99	97-102	103--108	97-114

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human SERPINE1 produced at Sunlong Medical™.

Ordering & shipping

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