DESCRIPTION

Tie2 (also known as TEK) is a tyrosine-protein kinase expressed almost exclusively on endothelial cells. It contains two immunoglobulin-like domains, three epidermal growth factor (EGF)-like domains and three fibronectin type III repeats. Tie2 acts as cell-surface receptor for ANGPT1, ANGPT2 and ANGPT4 and regulates angiogenesis, endothelial cell survival, proliferation, migration, adhesion and cell spreading, reorganization of the actin cytoskeleton, but also maintenance of vascular quiescence. Mutations in the gene of Tie2 are associated with inherited venous malformations of the skin and mucous membranes. Tie2 can also exist as a soluble form (sTie2) which is released from endothelial cells and present in human blood. The concentration of sTie2 is increased in a range of diseases, including peripheral arterial disease and myocardial infarction.

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	9.28	0.30	3.23
2	8	2.34	0.07	2.99
3	8	1.32	0.02	1.52

Inter-assay Precision

Sample	n	Mean (ng/mL)	SD	CV%
1	16	9.49	0.43	4.53
2	16	2.33	0.06	2.58
3	16	1.33	0.03	2.26

RECOVERY

The recovery of human Tie2 spiked to three different levels throughout the range of the assay in human serum was evaluated.

Sample Type		Average% of Expected	Range (%)
Human serum	1:32	112	109-117
	1:64	109	106-112

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 human serum was initially diluted 1:4.)

		Human serum
1:2	Average% of Expected	100
	Range (%)	-
1:4	Average% of Expected	107
	Range (%)	97-117
1:8	Average% of Expected	111
	Range (%)	92-129
1:16	Average% of Expected	104
	Range (%)	86-123

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human Tie2 produced at Sunlong Medical™.

Ordering & shipping

• Minimum order: 1 kit
• Lead time: varies with order quantity and specifications. Please consult your local distributor or our sales team for confirmation
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• Price: USD, per kit (48T / 96T options available)