- Fibrin zymography is a gel-based electrophoretic technique used to detect and analyze fibrinolytic enzymes—those that can degrade fibrin, the main protein component of blood clots.
- It is a sensitive and widely used method in both research and clinical studies to evaluate the activity of fibrinolytic enzymes in biological samples, microbial extracts, or purified proteins.
- Fibrin zymography is especially useful for detecting enzymes like plasmin, tPA, uPA, or microbial fibrinolytic enzymes such as streptokinase, nattokinase, and lumbrokinase. It allows researchers to determine both the presence and the approximate molecular weight of active enzymes, as well as to compare the activity levels between samples.
- This method is cost-effective, relatively simple, and provides visual, qualitative, and sometimes semi-quantitative information about fibrinolytic activity. It is often used alongside other biochemical assays to confirm enzyme identity and potency.
Procedure:
- The process involves running samples on a polyacrylamide gel that is co-polymerized with fibrin and plasminogen. During electrophoresis, proteins are separated based on their molecular weight under non-reducing and usually non-denaturing conditions.
- After electrophoresis, the gel is renatured (by removing SDS) and then incubated in a buffer that allows the fibrinolytic enzymes to regain their activity. Enzymes capable of degrading fibrin will do so at their specific position in the gel.
- After incubation, the gel is stained with Coomassie Brilliant Blue, which stains proteins, including the fibrin matrix. Clear bands against a blue background indicate areas where fibrin has been digested by active enzymes—these zones correspond to the molecular weights of the fibrinolytic enzymes present in the sample.
