Fibrinogen-PAGE for Fibrinogenolytic Agent Analysis in Peanut Worm Homogenate

To begin, prepare a gel for fibrinogen phage and place it in the electrophoresis tank together with the glue mold. Pour 1, 300 milliliters of electrophoresis solution into the tank and take out the comb from the gel. Mix 5 microliters of 5x non-denaturing loading buffer with 20 microliters each of fibrinogenolytic enzyme and peanut worm homogenate.

Load the mixture into the prepared gel and initiate the electrophoresis at 80 volts for 30 minutes or 120 volts for 20 minutes. Remove the gel from the glue mold and transfer it into a plastic box. Add Tris-HCL and Triton into the box and incubate it on a shaker at 60 to 70 RPM for 10 minutes.

Remove the wash buffer and incubate the gel with 50 milliliters of 0.05 molar Tris-HCL. After removing the buffer, incubate the gel with 50 milliliters of staining, followed by de-colorization solution at 60 to 70 RPM for 30 minutes each. The peanut worm homogenate displayed bands corresponding to molecular weights of 25, 33, 43, and 70 kilo-daltons or higher.