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我们用H~3-组氨酸掺入健康人全血嗜碱细胞,并观察其转化为~2H-组胺重新释放能力。 4名输血员,每份抗凝全血分成0.25ml一管,共4管,2管测掺入,2管测释放。分别加入0.1mlH~3-组氨酸(含5μCi)及50μl载体,37℃旋转温育20分钟,以冷Tyrode’s液洗涤3次,上清放射性本底可降至400opm以下。掺入管即可加0.1ml12.5%TAC,离心后吸上清待测(混合闪烁液,允许含水50μl/1ml)。释放管则加0.1mlConA(最终5μg/ml)及Ca~(++)、Mg~(++)继续温育50分钟,离心后吸上清测定。所有管均吸5μl在DEAE纤
We incorporated H ~ 3-histidine into healthy human whole blood basophils and observed their ability to be converted to ~ 2H-histamine re-release. Four blood transfusion, each anticoagulant whole blood is divided into a 0.25ml tube, a total of 4 tubes, 2 tubes measured incorporation, 2 tube test release. Add 0.1mlH3-histidine (containing 5μCi) and 50μl vector, incubate at 37 ° C for 20 minutes, wash with cold Tyrode’s solution three times, the supernatant radioactive background can be reduced to below 400opm. Addition tube can be added 0.1ml12.5% TAC, after centrifugation supernatant was measured (mixed scintillation fluid, allowing water 50μl / 1ml). Release tube and add 0.1mlConA (final 5μg / ml) and Ca ~ (++), Mg ~ (++) continue to incubate for 50 minutes, after centrifugation sucked supernatant determination. All tubes were pipetted 5 μl on DEAE Fibers