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 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. Recombinant TFPI-2 (Domain 1) produced by transient expression in non-transgenic plants includes a 6 His-tag at the N-terminal end and has a molecular weight of 9.3 kDa (79 amino acids residues). It is purified by sequential chromatography (FPLC) and does not contain any animal-derived components or impurities.
Human TFPI2 expressed in Nicotiana benthamiana
CAT# CSC-CTK0655-1 (1 mg); CAT# CSC-CTK0655-5 (5 mg)
Greater than 97% as determined by SDS-PAGE analysis.
The activity of the inhibitor is expressed as the amount of trypsin inhibited per milligram of inhibitor. The ability to prevent the hydrolysis of benzoyl-L-arginine ethyl ester hydrochloride by trypsin is measured by spectrophotometer. One mg protein will inhibit 1-1.5 mg trypsin with activity of approximately 10,000 BAEE units per mg protein.
Less than 0.04 EU/μg.
Lyophilized powder containing phosphate buffer salts (pH 7.1).
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. At higher concentration the solubility may be reduced and multimers generated. Soluble in water and in aqueous buffers of low ionic strengths. Repeated freeze-thaw cycles should be avoided.
Storage & Stability
The lyophilized protein, though stable at room temperature for up to 3 weeks, is best stored desiccated at -20°C. Reconstituted protein should be used immediately or stored long-term in undiluted working aliquots at -20°C. For long-term storage, it is recommended to add a carrier protein (0.1% HSA or BSA). Avoid repeated freeze-thaw cycles.
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