Synonyms: pancreatic trypsin inhibitor; trypsin inhibitor, pancreas type (bpti); trypsin-kallikrein inhibitor
Storage: 2-8°C
UNSPSC Code: 12352200
RIDADR: NONH for all modes of transport
Application:
Aprotinin is used for the protection of proteins and enzymes during isolation/purification. The inhibition of protease activity increases the lifetime of cells in cell and tissue culture studies.
• Further applications: Purification of urokinase, trypsin, and chymotrypsin on immobilized aprotinin
• Quantification of kallikrein activity in mixtures of esterases and proteases
• Controlled degradation of substrates by avoiding nonspecific proteolysis in clinical chemical tests
• Aprotinin as a model protein in protein-folding studies
• Molecular weight marker in SDS-polyacrylamide gel electrophoresis
General description:
Trypsin inhibitor, pancreas type from bovine lung. It is known as Pancreatic trypsin inhibitor (BPTI). Aprotinin, also known as pancreatic trypsin inhibitor and trypsin-kallikrein inhibitor, is found to be expressed in lungs, spleen, liver, and pancreas. It is also found to be present in the free form in calf serum.
Other Notes:
For life science research only. Not for use in diagnostic procedures.
Preparation Note:
Working concentration: 0.06 to 2 μg/ml (0.01 - 0.3 μM)
Working solution: Soluble in water (10 mg/ml) or aqueous buffer solution (e.g., 0.1 M Tris, pH 8.0).
Note: To avoid adsorption of aprotinin onto negatively charged solid phases, e.g., chromatography gels, ultrafiltration membranes, the NaCl concentration should be above 0.1 M or other suitable salts should be added to all buffers used during the separation.
Storage conditions (working solution): -15 to -25 °C
Reconstitution:
Freely soluble in water (10 mg/ml) or aqueous buffer solution (e.g., Tris, 0.1 M, pH 8.0). A solution adjusted to pH 7 to 8 is stable for approximately 1 week at 2 to 8 °C.
Aliquots stored at -15 to -25 °C are stable for approximately 6 months.
Note: Avoid repeated freezing and thawing and exposure to strongly alkaline solutions (inactive at pH > 12.8).
Sequence:
Monomeric peptide of 58 amino acids held in conformation by three disulfide bonds.
Specificity:
Aprotinin inhibits serine proteases. It inhibits kallikrein, the protease that releases hypotensive peptides such as kallidin and bradykinin (human plasma kallikrein: Ki = 3 ×10-8 M at pH 8.0, porcine pancreas kallikrein: Ki = 1 × 10-9 M at pH 8.0), trypsin (Ki = 2.8 × 10-11 M at pH 7.8, Ki = 2.6 × 10-9 M at pH 4.0, non-competetive), trypsinogen, chymotrypsin (Ki = 9 × 10-9 M at pH 8.0), bacterial fibrinolysin, and plasmin (Ki = 1 nM at pH 7.3).
Cathepsin G, acrosin, human leukocyte elastase, and human urokinase are weakly inhibited. Factor Xa, thrombin, subtilisin, papain, pepsin, angiotensin-converting enzyme (ACE), carboxypeptidase A and B, other metalloproteases, and thiolproteases are not inhibited.