从血浆中分离血清

Separating serum from plasma -- G (Blood plasma) - (fibrinogen & clotting factors) = Blood serum. 取决于抗凝剂：过量的氯化钙可使柠檬酸盐血浆（柠檬酸盐抗凝血，柠檬酸盐络合钙离 子）、EDTA 抗凝血或全血凝块(凝块收缩)，再次离心获得血清。仅从血浆分离血清时，Ca 的用量可按生理状态下血浆中的浓度计算或过量。 20% w/v CaCl2 into plasma to a ratio of 1:100 (v:v of CaCl2:plasma). Mix and lt clot at 4C overnight. Centrifuge to pellet down the clotting protein then you will get serum. 肝素抗凝血（heparinised blood），加过量的 thrombin or protamine sulphate（硫酸鱼精蛋 白），凝固后同上离心。protamine sulphate 的用量与 heparin 浓度相同，以中和 heparin 效应。 After clotting, centrifuge to get serum. Thrombin 的用量:You must add minimally 2 IU of thrombin to 1 ml of sample and after 2 minutes perform separation of clot by centrifugation at 4500 rpm minimally 5 min. If you allow the blood to clot you will get serum, not plasma. If you use an anticoagulant, you will get plasma, not serum. The reaction is not reversible since clotting removes all fibrinogen by converting it to fibrin. However, if you use heparin as an anticoagulant, it inhibits this conversion. If you remove heparin by using protamine sulphate, then coagulation will occur and is enhanced by the presence of calcium ions. 认为用 Ca 抗凝不可逆转，因为纤维蛋白原都转变成了纤维蛋白。而肝素抗凝可用硫酸 鱼精蛋白中和，此时加入 Ca 可促凝。 …use thrombin followed by calcium to convert plasma to serum (calcium alone - as calcium chloride, if I remember correctly - may do in some cases, depending on what was used as anticoagulant). I used to have a protocol but … and I can't find it. Once the clot has formed (leave it to shrink) you will need to centrifuge the serum + clot to get the serum layer, free of fibrin - or you could carry out the reaction by tumbling in a vessel with some glass beads which will attract the fibrin as they fall from one end of the vessel to the other when tumbled (we used to use a wheel rotator). This "tumbling on glass beads" can also be used for whole blood instead of anticoagulants - defibrinated blood is quite good if you want to isolate mononuclear leukocytes which are relatively free of platelets (which stick to the fibrin clot on the glass beads) - and you also get serum from the sample. 或者可以在管子中加入玻璃珠用颠倒法进行反应，玻璃珠可 吸附 fibrin（可以用旋转混合仪）。此法也可代替抗凝剂用于全血，去除纤维蛋白的血液用来 分离单核白细胞因血小板相对少效果很好（血小板粘附于玻璃珠上的纤维蛋白凝块），并且 可同时从其中获得血清。