Characterization of equine alkaline phosphatase isoenzymes based on their electrophoretic mobility by polyacrylamide gel disc electrophoresis.

The research paper focuses on the characterization of different types of equine alkaline phosphatase based on their movement in electric field via polyacrylamide gel disc electrophoresis. The characterization helps in understanding their origin in the tissues and possible contribution to serum and peritoneal fluid.

Methodology

• The researchers extracted alkaline phosphatase isoenzymes from various equine tissues including the liver, kidney, spleen, small intestine, placenta, bone, small colon, and large colon. These were then separated using a technique called polyacrylamide gel disc electrophoresis.
• Electrophoretic mobilities of the isolated isoenzymes were measured. These provide information about the size, shape and charge of these isoenzymes.
• A spectrophotometer was used to scan the separated isoenzymes. This helped in evaluating their tissue specificity.
• The researchers also tested the sensitivity of these isoenzymes to heat at 56 degrees Celsius. This may provide information about the stability of these molecules.

Findings

• The researchers were able to distinguish the isoenzymes obtained from all tissue extracts except those from small intestine and large colon based on their electrophoretic mobilities and heating sensitivities. This suggests that these isoenzymes have similar sizes, shapes, charges and stability.
• However, the heat resistance characteristic of the isoenzyme extracted from the small intestine was unique enough to distinguish it from the large colon.
• The specific mobilities of isoenzymes helped to determine the possible contribution of alkaline phosphatase in serum and peritoneal fluid.

Applications

• This study could prove significant in equine medicine by helping to diagnose diseases associated with these tissues such as liver disease or bone disorders which may involve an abnormal amount of alkaline phosphatase in the blood or peritoneal fluid.
• It can also be used to monitor the health status of pregnant mares and foals by evaluating the isoenzyme patterns in the serum.