DESCRIPTION

Thrombospondin 1(TSP-1), an extracellular matrix protein, is the first identified natural angiogenesis inhibitor. A variety of normal cells, including endothelial cells, fibroblasts, adipocytes, smooth muscle cells, monocytes, macrophages, and transformed cells such as malignant glioma cells, secrete Thrombospondin 1. It regulates cellular phenotype during tissue genesis and repair. It acts as a molecular facilitator by bringing together cytokines, growth factors, matrix components, membrane receptors and extracellular proteases. Thrombospondin 1 binds to a wide variety of integrin and non-integrin cell surface receptors. It is also a major activator of transforming growth factor (TGFβ1).

PRINCIPLE OF THE ASSAY

Sandwich ELISA structure (Detection antibody labeled with biotin) A capture antibody is pre-coated onto the bottom of wells which binds to analyte of interest. A detection antibody labeled with biotin also binds to the analyte. Streptavidin-HRP binds to the biotin. 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	44.43	4.17	9.4
2	20	3.71	0.31	8.3
3	20	0.31	0.03	9.6

Inter-assay Precision

Sample	n	Mean (ng/mL)	SD	CV%
1	24	36.57	3.67	10.0
2	24	3.66	0.20	5.5
3	24	0.34	0.03	8.3

RECOVERY

The recovery of human Thrombospondin 1 spiked to three different levels throughout the range of the assay in various matrices was evaluated.

Sample Type		Average% of Expected	Range (%)
Human serum	1:3000	84	75-101
	1:6000	78	70-84
Cell culture supernatant	1:300	87	72-100
	1:600	88	83-96
saliva	1:30	80	70-101
	1:60	88	77-117
Human milk	1:100	95	83-114
	1:200	81	77-93

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 human serum were initially diluted 1:800, the cell culture supernatant was initially diluted 1:120, the saliva were initially diluted 1:8, the human milk were initially diluted 1:8)

		Human serum	Cell culturesupernatant	Saliva	Human milk
1:2	Average% ofExpected	100	100	100	100
	Range (%)	-	-	-	-
1:4	Average% ofExpected	107	103	107	99
	Range (%)	99-114	94-112	101-112	96-102
1:8	Average% ofExpected	113	113	101	95
	Range (%)	105-123	111-115	100-103	94-98
1:16	Average% ofExpected	113	121	112	104
	Range (%)	104-123	115-124	106-117	97-116

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human Thrombospondin 1 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)