Fontaine N, Meslin J C
INRA, laboratoire de nutrition et sécurité alimentaire, Jouy-en-Josas, France.
Reprod Nutr Dev. 1994;34(3):237-47.
A method, transposed from the mucin histochemical stainings, was proposed to evaluate neutral, acidic and sulphated mucins by spectrocolorimetry. Stainings used were periodic acid/Schiff (PAS), Alcian blue (AB) pH 2.5, and Alcian blue (AB) pH 0.5. Mucin samples were extracted from mucosal scrapings, from intestinal contents of germ-free rats or from commercial pig gastric mucin. A mucin-enriched fraction was obtained and lyophilised and the protein content was determined. Each mucin type was spectrophotometrically analysed after staining and precipitation by slightly modified Carnoy fixative. The histochemical stainings used here, after modification by Carnoy fixative treatment, were, as shown by electrophoretic controls, specific for mucin types contained in these mucins without protein contaminant interference.
提出了一种从粘蛋白组织化学染色方法转换而来的方法,通过分光比色法评估中性、酸性和硫酸化粘蛋白。使用的染色方法为过碘酸/希夫(PAS)、pH 2.5的阿尔辛蓝(AB)和pH 0.5的阿尔辛蓝(AB)。粘蛋白样本从黏膜刮片、无菌大鼠的肠道内容物或市售猪胃粘蛋白中提取。获得富含粘蛋白的部分并冻干,然后测定蛋白质含量。每种粘蛋白类型在经轻微改良的卡诺固定剂染色和沉淀后进行分光光度分析。如电泳对照所示,此处使用的经卡诺固定剂处理改良后的组织化学染色,对这些粘蛋白中所含的粘蛋白类型具有特异性,且无蛋白质污染物干扰。