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Corn Trypsin Inhibitor (CTI)


Effect of CTI on the APTT and PT of Normal Human Plasma
The graph illustrates the effect of added CTI on the PT and aPTT of normal human plasma. CTI was added to normal pooled human plasma to various final concentrations, and the mixtures were allowed to incubate for a brief period on ice prior to conducting the assays. The PT remains unaffected while the aPTT doubles with the addition of approximately 60 micrograms of CTI per ml of plasma.

  • Price $90.00/1mg ($85.00/min. 10)
    Size 1mg
    Formulation 20 mM Tris, 150 mM NaCl, pH 7.4
    Molecular Weight
    Storage -80°C
    Purity >95% by SDS-PAGE
    Compound
    Assay aPTT assay
    Shelf Life (properly stored) 12 months
    Chemical Formula
  • Prices $338.00/25 X 2 mL , $468.00/25 X 3 mL , $524.25/25 X 5 mL
    Sizes 25 X 2 mL, 25 X 3 mL, 25 X 5 mL
    Formulation 11 mM Citrate, 50 ug/mL CTI (final)
    Molecular Weight
    Storage 4°C
    Purity N/A
    Compound
    Assay N/A
    Shelf Life (properly stored) 6 months
    Chemical Formula
Gel Novex 4-12% Bis-Tris
Load Corn Trypsin Inhibitor, 1 µg per lane
Buffer MES
Standard SeeBluePlus 2; Myosin (188 kDa), Phosphorylase B (98 kDa), BSA (62 kDa), Glutamic Dehydrogenase (49 kDa), Alcohol Dehydrogenase (38 kDa), Carbonic Anhydrase (28 kDa), Myoglobin Red (17 kDa), Lysozyme (14 kDa), Aprotinin (6 kDa), Insulin, B chain (3 kDa).

Purified CTI
Corn trypsin inhibitor (CTI) is a small protein that is localized in the kernels of most species of corn. CTI is not only an inhibitor of trypsin, but is also a specific human factor XIIa inhibitor when tested in blood clotting experiments (1-6). The inhibitor forms a one-to-one complex with either trypsin or factor XIIa, and when added to plasma, prolongs the activated partial thromboplastin time without affecting the PT assay (6). The specificity for factor XIIa makes the inhibitor useful for the segregation and study of tissue factor (TF) dependent coagulation reactions (6).

The use of CTI to study TF-dependent reactions has been documented in literature (6, 8-10). Studies indicate that suppression of the contact pathway of coagulation is essential when attempting to perform TF-dependent assays in whole blood or plasma samples. The addition of CTI at the point of sample collection prevents activation of the contact pathway during subsequent sample processing steps, thus reducing in-vitro artifact. The most common use of CTI is associated with thrombin generation assays when attempting to work at low TF concentrations. HTI’s CTI product is performance tested for this specific application.

The protein is comprised of 112 amino acids which yields a calculated molecular weight of 12,028 (7). The calculated molecular weight and that which has been determined by sedimentation equilibrium analyses of the purified protein (12,500) are in good agreement (3). A comparison of the amino acid sequence of CTI to that of other trypsin or serine protease inhibitors reveals little or no similarities (7).

CTI is purified using a combination of published procedures (3,4). To begin the purification process, CTI is extracted from the kernels of fresh sweet corn into a physiologic buffer. The extract is then de-fatted using acetone, and the protein is further purified by employing gel filtration and ion-exchange chromatography. The final CTI preparation appears as a single band by SDS-PAGE analyses under both reducing and non-reducing conditions.

Preparations of CTI are tested for the ability to prolong the aPTT assay without affecting the PT assay. The specific activity of each lot of CTI is determined, and one unit is defined as the amount of CTI required to double the aPTT of normal human plasma.

Purified CTI is formulated in 20 mM Tris, 0.15 M NaCl, pH 7.4 and should be stored frozen at -20°C or colder.

Blood Collection Tubes
To simplify the process of collecting samples with added CTI, HTI has developed blood collection tubes containing CTI. These tubes (our SCAT-27 line) simplify the process of conducting TF-dependent studies by allowing you to draw blood directly onto an anticoagulant containing CTI. You may choose to use our standard CTI/Citrate formulation (11mM Citrate, 50 ug/mL CTI) or you may create your own custom formulation. HTI’s blood collection tubes are not sterile and are manufactured and sold for research use only. Three standard sizes are available although custom sizes can be manufactured for you.

SCAT-27-4.5/5 - 5 mL tube

SCAT-27-2.7/5 - 3 mL tube

SCAT-27-1.8/5 - 2 mL tube

 

Localization Corn Kernals
Mode of action forms a one-to-one complex with either trypsin or factor XIIa and inhibits their respective catalytic activity.
Molecular weight 12,500 (3)
Extinction coefficient
E
1 %
1 c m, 280 nm
= 20.0 (determined by ultra centrifugation studies)
Structure single chain protein comprised of 112 amino acids (3,7)
  1. Hochstrasser, K., et al., Z. Physiol. Chem., 348, 1337 (1967).
  2. Halim, R.K., et al., Crop. Sci., 13, 405 (1973).
  3. Swartz, M.J., et al., J. Biol. Chem., 252, 8105 (1977).
  4. Hojima, Y., et al., Thromb. Res., 20, 149 (1980).
  5. Ratnoff, O.D. and Moneme, V., Proc. Soc. Exp. Biol. Med., 166, 297 (1981).
  6. Rand, M.D. et al., Blood, 88, 3432 (1996).
  7. Mahoney, W.C., et al., J. Biol. Chem., 259, 8412 (1984).
  8. Cawthern KM., et al., Blood. 1998;91:4581-4592.
  9. Dargaud Y, et al., Journal of Thrombosis and Haemostasis. 2006;4:1160-1161.
  10. Mann K.G., et al., J Thromb Haemost. 2007;5:2055-2061.
  1. Pedicord, D., et al., Proc Natl Acad Sci U S A. 2007 July 31; 104(31): 12855–12860. (CTI in collection tubes)
  2. Buyue, Y., et al., 2008 112: 3234-3241 (Thrombin generation)
  3. Rivard, G., et al., J Thromb Haemost. 2005 September ; 3(9): 2039–2043. (Thrombin generation)
  4. Okorie, U., et al, Biophys J. 2006 November 1; 91(9): 3474–3481. (inhibition of intrinsic pathway)

This publication list is not all encompassing, and is only meant to provide limited examples of how Haematologic Technologies' products are used. We encourage you to search the literature for other examples pertinent to your experimentation, and to contact us with any technical questions.