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Purification of Mouse Brain Vessels
Published on: November 10, 2015
Purification of human brain tissue factor
Thrombosis and Haemostasis
|June 16, 1988
Summary
Researchers purified human brain tissue factor (Factor III), essential for blood coagulation, achieving 98% homogeneity. This advancement utilized affinity chromatography for efficient isolation of this crucial glycoprotein cofactor.
Area of Science:
- Biochemistry
- Hematology
- Molecular Biology
Background:
- Tissue factor (Factor III) is a critical glycoprotein cofactor.
- It is essential for initiating both extrinsic and intrinsic blood coagulation pathways.
- Factor VII requires tissue factor for its catalytic activity.
Purpose of the Study:
- To develop an efficient purification method for human brain tissue factor.
- To achieve high purity and yield of the tissue factor.
- To characterize the purified tissue factor.
Main Methods:
- Purification of human brain tissue factor from acetone-dried brain powder using Triton X-100 extraction.
- Affinity chromatography employing immobilized anti-Factor VII-agarose beads.
- Characterization using SDS-PAGE and isoelectric focusing techniques.
Main Results:
- Achieved a 2,500-fold purification of tissue factor to 98% homogeneity.
- Obtained an overall yield of 36% for the purified tissue factor.
- Determined the apparent molecular weight to be 45,000 Da and isoelectric point to be 4.8-5.1.
Conclusions:
- A robust method for purifying human brain tissue factor was established.
- The purified tissue factor is highly homogeneous and characterized.
- This facilitates further research into the role of tissue factor in coagulation.
