| Cat. No. : |
TFPI2-008N |
| Description : |
AGV 212 is a protease inhibitor peptide generated from the first Kunitz domain of the human Tissue Factor Protein Inhibitor 2 (TFPI-2) protein, after site-directed mutagenesis to increase its activity. It consists in 79 amino acids (including 6 His located in the N-terminus end, for tagging purposes). It is arranged in a single polypeptide chain that is linked by three disulfide bridges. AGV 212 is quite stable and inhibits trypsin with high efficiency and Ki lower than TFPI-2 one. TFPI-2 has been shown to inhibit Endothelial Cell Matrix (ECM) proteases essential for angiogenesis and metastasis. This product contains no animal or bacterial-derived components or impurities. It is manufactured by transient expression in non-transgenic plants. |
| Source : |
Nicotiana benthamiana. |
| Formulation : |
Lyophilized powder containing phosphate buffer salts, pH 7.1. |
| Molecular formula : |
C407H592N116O117S6 |
| Extinction coefficient : |
E0.1% = 2.35 (A280 nm). |
| Molecular weight : |
9.174 Da |
| Purity assay : |
≥ 97% (SDS-PAGE) |
| Endotoxin level : |
< 0.04 EU/ µg protein (LAL method) |
| Reconstitution recommendation : |
The lyophilized protein should be reconstituted adding 1 ml of sterile water to the vial, which gives a concentration of 1 mg of protease inhibitor per ml. In higher concentration the solubility may be reduced and multimers generated. AGV 212 is soluble in water and in aqueous buffer of low ionic strengths. Repeated freeze-thaw cycles should be avoided. |
| Biological Activity : |
The ability of the trypsin inhibitor AGV 212 to prevent trypsin hydrolysis of benzoyl-L-arginine ethyl ester hydrochloride (B4500 Sigma) is measured by spectrophotometer. The activity of the inhibitor is expressed as the amount of trypsin (T8003 Sigma) inhibited by one milligram of inhibitor. Operating procedure Sigma Aldrich # 10-30-6920 One trypsin unit = ?A253 of 0.001 per min with BAEE as substrate at pH 7.6 at 25°C. Reaction volume = 3.2 ml (1 cm light path). One mg protein will inhibit 1-1.5 mg trypsin with activity of approximately 10,000 BAEE units per mg protein. |
| Storage : |
This lyophilized preparation is stable at 2-8℃, but should be kept at -20℃ for long term storage. The product, as supplied is stable for at least 2 years. Diluted solutions are less stable than concentrated ones. Repeated freezing and thawing is not recommended. |
