Aprotinin, also called bovine pancreatic trypsin inhibitor, is a single chain globular Kunitz-type serine proteinase inhibitor found in bovine pancreas, parotid gland and lung. It forms stable, enzymatically inactive complexes with trypsin, chymotrypsin, plasmin and kallikrein. Aprotinin has been used therapeutically as an antifibrinolytic agent under the name Trasylol (Bayer) to reduce blood loss during cardiopulmonary bypass surgery but this use has been associated with an increased frequency of acute renal injury (1).
Applications:
Suitable for use in ELISA. Other applications not tested.
Recommended Dilution:
ELISA: 1:33,000. 2ug/ml of A2300-01C forms an antibody pair with A2300-01B (as biotinylated detection antibody) for sandwich ELISA of aprotinin at sample concentrations of 0.6-20ug/ml. Aprotinin in urine samples diluted 1:1000 can be assayed against standards in buffer. For assay in plasma, it is recommended to add aprotinin-free control plasma to the standards to match the plasma sample dilution. PBS-based buffers are unsuitable for this assay and 0.2M glycine sodium buffer, pH 8.8, is recommended for washing and dilution buffers.
Optimal dilutions to be determined by the researcher.
Culture Medium: RMPI 1640 with 2-10% fetal calf serum.
Hybridoma:
Sp2mIL6 myeloma cells with spleen cells from NMRI x Balb/c mice.
Storage and Stability:
May be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
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