Munakata H, Isemura M, Yosizawa Z
Tohoku J Exp Med. 1985 Mar;145(3):251-7. doi: 10.1620/tjem.145.251.
The high-iron diamine staining (HID), which has been used in histochemistry to stain sulfated glycoconjugates (SGC), was tested for detectability of authentic acidic substances (chondroitin sulfates A plus C, dermatan sulfate, heparan sulfate, chondroitin, hyaluronic acid, alpha 1-acid glycoprotein and ribonucleic acid) in electrophoresis on cellulose acetate membrane (Separax). The results showed that only SGC were detectable by the HID, although all the above substances were stained with alcian blue. The glycoconjugate preparations obtained from the liver, kidney, lung, small intestine, colon, stomach, brain and spleen of rats were examined by two-dimensional electrophoresis on Separax. The new spots (or bands), besides those of sulfated glycosaminoglycans, were detected by the HID on the electrophoretograms of all the samples except for the kidney one. The substances giving the new spots (or bands) were indicated to be sulfated glycopeptides (SGP) by crude heparinase digestion of a representative sample. The present results revealed that the HID was applicable for detection of SGP in electrophoresis on cellulose acetate membrane. Also, it is a novel finding that the liver and spleen contain SGP.
高铁二胺染色法（HID）曾用于组织化学中对硫酸化糖缀合物（SGC）进行染色，本研究对其在醋酸纤维素膜（Separax）电泳中检测真实酸性物质（硫酸软骨素A加C、硫酸皮肤素、硫酸乙酰肝素、软骨素、透明质酸、α1-酸性糖蛋白和核糖核酸）的能力进行了测试。结果表明，尽管上述所有物质均能用阿尔辛蓝染色，但HID只能检测到SGC。对从大鼠的肝脏、肾脏、肺、小肠、结肠、胃、脑和脾脏中获得的糖缀合物制剂在Separax上进行二维电泳分析。除肾脏样本外，在所有样本的电泳图谱上，HID均检测到了除硫酸化糖胺聚糖斑点（或条带）之外的新斑点（或条带）。通过对一个代表性样本进行粗制肝素酶消化，发现产生这些新斑点（或条带）的物质为硫酸化糖肽（SGP）。本研究结果表明，HID可用于在醋酸纤维素膜电泳中检测SGP。此外，肝脏和脾脏中含有SGP这一发现尚属首次。
