Briggs R C, Glass W F, Montiel M M, Hnilica L S
J Histochem Cytochem. 1981 Oct;29(10):1128-36. doi: 10.1177/29.10.7028857.
Conditions were established whereby nuclear or cytoplasmic immunocytochemical localization of lactoferrin was observed in the human peripheral blood granulocyte. A positive nuclear staining reaction was obtained when cells were either not fixed or treated with most fixatives. However, treatment of blood smears with formaldehydeacetone prior to the immunocytochemical localization gave a cytoplasmic staining reaction. A method was developed that allowed us to examine proteins obtained from granulocyte nuclei isolated from fixed cells. Only a trace amount of lactoferrin was detected in the electrophoretically separated nuclear proteins obtained from granulocytes treated for 1 min with formaldehyde-acetone. However, lactoferrin was major protein component in nuclei isolated from untreated cells, cells treated with other fixatives, or cells preincubated in buffered saline prior to formaldehyde-acetone fixation. formaldehyde-acetone treatment of nuclear materials containing lactoferrin did not extract lactoferrin or mask it from detection, thus indicating that lactoferrin found in the mature neutrophilic granulocyte nucleus is most likely acquired from other cellular organelles during tissue processing.
建立了在人外周血粒细胞中观察乳铁蛋白核或细胞质免疫细胞化学定位的条件。当细胞不固定或用大多数固定剂处理时,可获得阳性核染色反应。然而,在免疫细胞化学定位之前用甲醛 - 丙酮处理血涂片会产生细胞质染色反应。开发了一种方法,使我们能够检测从固定细胞中分离的粒细胞核中获得的蛋白质。在用甲醛 - 丙酮处理1分钟的粒细胞的电泳分离核蛋白中仅检测到微量乳铁蛋白。然而,乳铁蛋白是从未处理细胞、用其他固定剂处理的细胞或在甲醛 - 丙酮固定之前在缓冲盐水中预孵育的细胞中分离的核中的主要蛋白质成分。对含有乳铁蛋白的核材料进行甲醛 - 丙酮处理不会提取乳铁蛋白或使其无法检测到,因此表明在成熟嗜中性粒细胞核中发现的乳铁蛋白很可能是在组织处理过程中从其他细胞器获得的。