免疫组织化学中在不使抗原变性的情况下对内源性过氧化物酶的有效抑制。

Efficient inhibition of endogenous peroxidase without antigen denaturation in immunohistochemistry.

作者信息

Malorny U, Bildau H, Sorg C

机构信息

Institute of Experimental Dermatology, University of Münster, F.R.G.

出版信息

J Immunol Methods. 1988 Jun 28;111(1):101-7. doi: 10.1016/0022-1759(88)90065-8.

DOI:10.1016/0022-1759(88)90065-8

PMID:3292649

Abstract

The immunoperoxidase technique is commonly used for the antigenic characterization of cells and tissues. However, the occurrence of endogenous peroxidase-positive cells frequently complicates such evaluations. In some instances it is impossible to distinguish between these cells and antibody-stained cells. Methods so far described for inhibiting the endogenous peroxidase, are not always satisfactory - either the endogenous peroxidase is not satisfactorily blocked, or the cellular antigens lose their reactivity. A new two-step method is described which involves transient inhibition of endogenous peroxidase by sodium azide and hydrogen peroxide and a final blocking of this enzyme by lowering the pH to 2.3, while antigens remain undamaged. This method may be of value in immunohistochemical studies of various tissues, especially inflammatory tissues which contain many endogenous peroxidase-positive cells.

摘要

免疫过氧化物酶技术常用于细胞和组织的抗原特性鉴定。然而，内源性过氧化物酶阳性细胞的存在常常使此类评估变得复杂。在某些情况下，无法区分这些细胞与抗体染色的细胞。迄今为止所描述的抑制内源性过氧化物酶的方法并不总是令人满意——要么内源性过氧化物酶未被充分阻断，要么细胞抗原失去其反应性。本文描述了一种新的两步法，该方法包括用叠氮化钠和过氧化氢短暂抑制内源性过氧化物酶，然后通过将pH值降至2.3来最终阻断该酶，同时抗原保持未受损状态。该方法在各种组织的免疫组织化学研究中可能具有价值，尤其是含有许多内源性过氧化物酶阳性细胞的炎症组织。