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DACM[N-(7-dimethylamino-4-methylcou-marinyl)-maleimide]可作为一种敏感的荧光标记物。实验证明 DACM 标记兔抗人 IgG 的结合物可用葡聚糖凝胶 G50胶滤法排除未反应物,而且此标记反应不影响抗体的免疫学活性。标记方法将兔抗人 IgG 免疫血清按一般方法提取并精制 IgG,用 pH 8.2的0.55M Tris-HCl 缓冲液配成8mg IgG/ml 溶液,于20℃用抽气法排气1小时。加入2-巯基乙醇(2-ME)到最终浓度为4mM,于20℃静置1小时。将 DACM 丙酮溶液加入到 IgG 液中使其浓度达4mM,于20℃静置1小时。在pH 7.4的5mM PBS 中透析过夜,通过葡
DACM [N- (7-dimethylamino-4-methylcou-marinyl) -maleimide] can be used as a sensitive fluorescent label. Experiments show that DACM labeled rabbit anti-human IgG conjugates can be gelled with dextran G50 gel filtration unreacted material, and this labeling reaction does not affect the immunological activity of the antibody. Labeling Method Rabbit anti-human IgG serum was extracted and purified according to a general method. The solution was diluted to 8 mg IgG / ml with 0.55 M Tris-HCl buffer, pH 8.2, and degassed by suction at 20 ° C for 1 hour. 2-Mercaptoethanol (2-ME) was added to a final concentration of 4 mM and allowed to stand at 20 ° C for 1 hour. The DACM acetone solution was added to the IgG solution to a concentration of 4 mM and allowed to stand at 20 ° C for 1 hour. Dialyzed overnight in 5 mM PBS, pH 7.4, through the port of glucose