使用二氨基联苯胺（DAB）在红细胞、中性粒细胞和过氧化物酶-抗过氧化物酶复合物中定位过氧化物酶的方法比较。

A comparison of methods using diaminobenzidine (DAB) to localize peroxidases in erythrocytes, neutrophils, and peroxidase-antiperoxidase complex.

作者信息

Vacca L L, Hewett D, Woodson G

出版信息

Stain Technol. 1978 Nov;53(6):331-6. doi: 10.3109/10520297809111955.

DOI:10.3109/10520297809111955

Abstract

Reactions using diaminobenzidine (DAB) to localize the enzyme peroxidase in neutrophils and peroxidase-antiperoxidase (PAP) complex during immunological staining are usually performed in Tris-HCl or phosphate buffer at pH 7.2-7.6. However, DAB solutions at pH 7.2-7.6 often demonstrate erythrocyte pseudoperoxidase as well. By lowering the pH of the DAB solutions, it is possible to selectively suppress the reactivity of pseudoperoxidase while maintaining optimal reactions in neutrophils and PAP complex. For this purpose we recommend ammonium acetate-citric acid buffer at pH 5.5 (pH 5.0-6.0) containing 44 mg DAB per 100 ml buffer and 0.003%-0.03% with respect to H2O2.

摘要

在免疫染色过程中，使用二氨基联苯胺（DAB）对中性粒细胞中的过氧化物酶和过氧化物酶-抗过氧化物酶（PAP）复合物进行定位的反应，通常在pH 7.2 - 7.6的Tris-HCl或磷酸盐缓冲液中进行。然而，pH 7.2 - 7.6的DAB溶液也常常显示出红细胞假过氧化物酶活性。通过降低DAB溶液的pH值，可以在保持中性粒细胞和PAP复合物中最佳反应的同时，选择性地抑制假过氧化物酶的反应活性。为此，我们推荐使用pH 5.5（pH 5.0 - 6.0）的醋酸铵-柠檬酸缓冲液，每100 ml缓冲液中含有44 mg DAB，相对于H2O2的浓度为0.003% - 0.03%。