DESCRIPTION

Plasminogen activator, tissue (PLAT, synonyms: TPA, T-PA) is a tissue-type plasminogen activator, a secreted serine protease which converts the proenzyme plasminogen to plasmin, a fibrinolytic enzyme. PLAT enzyme plays a role in cell migration and tissue remodeling. Increased enzymatic activity causes hyperfibrinolysis, which manifests as excessive bleeding; decreased activity leads to hypofibrinolysis which can result in thrombosis or embolism.

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	21.51	0.56	2.60
2	8	5.23	0.16	3.06
3	8	2.44	0.08	3.28

Inter-assay Precision

Sample	n	Mean (ng/mL)	SD	CV%
1	16	21.96	0.76	3.46
2	16	5.27	0.16	3.04
3	16	2.48	0.10	4.03

RECOVERY

The recovery of mouse t-Plasminogen activator spiked to three different levels throughout the range of the assay in mouse serum was evaluated.

Sample Type		Average% of Expected	Range (%)
Mouse serum	1:8	111	108-116
	1:16	108	101-116

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.

		Mouse serum
1:2	Average% of Expected	100
	Range (%)	-
1:4	Average% of Expected	112
	Range (%)	109-115
1:8	Average% of Expected	109
	Range (%)	105-113
1:16	Average% of Expected	112
	Range (%)	105-119

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant mouse t-Plasminogen activator produced at Sunlong Medical™.