Imam S A, Young L, Chaiwun B, Taylor C R
Department of Pathology, University of Southern California School of Medicine, Los Angeles 90033, USA.
Anticancer Res. 1995 Jul-Aug;15(4):1153-8.
The present study compared two microwave based antigen-retrieval solutions in their ability to unmask antigenic determinants in formalin-fixed and paraffin-embedded tissues for Immunostaining. In this regard, two widely used antigen-retrieval solutions, namely 0.05 M glycine-HCl buffer, pH 3.6, containing 0.01% (w/v) (EDTA) and 0.1 M sodium citrate buffer, pH 6.0, were evaluated for (1) their effectiveness in unmasking a wide range of antigenic determinants (2) their ability to yield reproducible results (3) the lack of deleterious effects in any antibody antigen systems of interest. Both of these antigen-retrieval solutions resulted in greatly improved immunostaining following microwave-heating of dewaxed tissue sections for 2 x 5 min. Glycine-HCl buffer solution resulted in stronger immunostaining with antibodies to nuclear antigens [androgen receptor (AR), estrogen receptor (ER), progesterone receptor (PR), p53, proliferating cell nuclear antigen (PCNA), Ki-67 and MIB-1], cytoplasmic antigens (actin and factor-VIII) and cell-surface antigens [Cu-18, epithelial membrane antigen (EMA) and MT-1 (CD43)], whereas sodium citrate buffer yielded superior immunostaining with antibodies to vimentin, and some cell-surface antigens [common leukocyte antigen (CLA) (CD45) and UCHL-1 (CD45RO)]. The effect of unmasking the epitopes recognized by antibody to PCNA was equally effective with either of the antigen-retrieval solutions. Antibodies to pan-keratin, prostatic acid phosphatase (PAP), B lymphocyte antigen (BLA.36, CD20CY) and L26 (CD20) exhibited no enhancement in the intensity of staining with either of the antigen-retrieval solutions.
本研究比较了两种基于微波的抗原修复溶液在福尔马林固定石蜡包埋组织中用于免疫染色时，使抗原决定簇暴露的能力。在这方面，对两种广泛使用的抗原修复溶液进行了评估，即pH 3.6的0.05 M甘氨酸 - HCl缓冲液（含0.01%（w/v）乙二胺四乙酸（EDTA））和pH 6.0的0.1 M柠檬酸钠缓冲液，评估内容包括：（1）它们在暴露多种抗原决定簇方面的有效性；（2）产生可重复结果的能力；（3）在任何感兴趣的抗体 - 抗原系统中均无有害影响。将脱蜡后的组织切片经微波加热2×5分钟后，这两种抗原修复溶液均使免疫染色得到显著改善。甘氨酸 - HCl缓冲液在用针对核抗原（雄激素受体（AR）、雌激素受体（ER）、孕激素受体（PR）、p53、增殖细胞核抗原（PCNA）、Ki - 67和MIB - 1）、细胞质抗原（肌动蛋白和因子VIII）以及细胞表面抗原（Cu - 18、上皮膜抗原（EMA）和MT - 1（CD43））的抗体进行免疫染色时，能产生更强的染色效果；而柠檬酸钠缓冲液在用针对波形蛋白以及某些细胞表面抗原（共同白细胞抗原（CLA）（CD45）和UCHL - 1（CD45RO））的抗体进行免疫染色时，能产生更好的染色效果。两种抗原修复溶液在使PCNA抗体识别的表位暴露方面效果相同。针对泛角蛋白、前列腺酸性磷酸酶（PAP）、B淋巴细胞抗原（BLA.36，CD20CY）和L26（CD20）的抗体在用任何一种抗原修复溶液染色时，染色强度均未增强。
