摘要
A technique was developed for detecting venom phospholipase A2(PLA2) using serum and red blood cells. M1 and M2, two PLA2s isolated from Crotalus m.molossus (Northern black-tail rattlesnake), were used for preliminary development of the as say. Various combinations of human, sheep, rat, and mouse red blood cells (RBC) with human,rat,and mouse sera were tested on their effectiveness to detect PLA2.Complete hemolysis (b hemolysis) was evident in the plate with rat RBC mixed with mouse serum.No hemolysis was detected in plates containing human RBC and human serum. Human RBC mixed with mouse serum proved to be ideal, even though this combination displayed incomplete hemolysis(a hemolysis).Susceptible RBC, in conjunction with rat or mouse serum, are excellent indicators for the presence of PLA2.Mixtures of RBC and serum in combination with BB4(E. coli) cells,λbacteriophage,and IPTG on LB agar plates provide an excellent detection system for cDNA clones that express venonl PLA2. Hemolysis surrounding a plaque is identified as positive for PLA2.
A technique was developed for detecting venom phospholipase A2(PLA2) using serum and red blood cells. M1 and M2, two PLA2s isolated from Crotalus m.molossus (Northern black-tail rattlesnake), were used for preliminary development of the as say. Various combinations of human, sheep, rat, and mouse red blood cells (RBC) with human,rat,and mouse sera were tested on their effectiveness to detect PLA2.Complete hemolysis (b hemolysis) was evident in the plate with rat RBC mixed with mouse serum.No hemolysis was detected in plates containing human RBC and human serum. Human RBC mixed with mouse serum proved to be ideal, even though this combination displayed incomplete hemolysis(a hemolysis).Susceptible RBC, in conjunction with rat or mouse serum, are excellent indicators for the presence of PLA2.Mixtures of RBC and serum in combination with BB4(E. coli) cells,λbacteriophage,and IPTG on LB agar plates provide an excellent detection system for cDNA clones that express venonl PLA2. Hemolysis surrounding a plaque is identified as positive for PLA2.
