Dey P, Saphos C A, McDonnell J, Moore V L
Merck Research Laboratories, Rahway, New Jersey.
Connect Tissue Res. 1992;28(4):317-24. doi: 10.3109/03008209209016823.
The dimethylmethylene blue (DMMB) dye-binding technique is widely used for the quantification of sulfated glycosaminoglycans (sGAG) and proteoglycans. We conducted further studies on this technique in our laboratory and found that concentrations of DNA and RNA in excess of 20 micrograms/ml interfered negatively with the detection of sGAGs; interference was eliminated by using DNase and RNase. Hyaluronan at 40 micrograms per ml did not interfere with the detection of sGAG. However, because of the higher concentrations of hyaluronan in synovial lavage fluid, it was necessary to treat this fluid with Streptomyces hyaluronidase in order to quantify sGAG. The DMMB assay was automated with a laboratory work station and compared to the standard method.
二甲基亚甲基蓝（DMMB）染料结合技术被广泛用于硫酸化糖胺聚糖（sGAG）和蛋白聚糖的定量分析。我们在实验室对该技术进行了进一步研究，发现DNA和RNA浓度超过20微克/毫升会对sGAG的检测产生负面影响；使用脱氧核糖核酸酶（DNase）和核糖核酸酶（RNase）可消除干扰。每毫升40微克的透明质酸不会干扰sGAG的检测。然而，由于滑膜灌洗液中透明质酸浓度较高，为了对sGAG进行定量分析，有必要用透明质酸酶链霉菌处理该液体。使用实验室工作站实现了DMMB分析的自动化，并与标准方法进行了比较。
