A RAPID METHOD FOR PREPARATION OF ROUGH AND SMOOTH LIPOPOLYSACCHARIDE FROM BACTEROIDES, PORPHYROMONAS AND PREVOTELLA
A RAPID METHOD FOR PREPARATION OF ROUGH AND SMOOTH LIPOPOLYSACCHARIDE FROM BACTEROIDES, PORPHYROMONAS AND PREVOTELLA
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DOI：
10.1016/0378-1097(93)90455-b
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发表时间：
1993-06-15
影响因子：
2.1
通讯作者：
MOUTON, C
中科院分区：
文献类型：
--
作者：
EIDHIN, DN;MOUTON, C
本文介绍了一种新的制备脂多糖的方法，即在100℃下用水提取，然后用蛋白酶K消化，粗制的脂多糖保留了很高的抗原性，不含十二烷基硫酸钠和蛋白酶K，是一种可靠的免疫印迹方法。用两种常规方法制备的两株不能与内毒素反应的单抗与我们的制备方法反应良好。我们使用这种新方法从44株以前被归类为类杆菌的细菌中提取内毒素，其中一些已被重新归类为卟啉单胞菌或普雷沃特氏菌。总体而言，产量是好的，通过SDS-PAGE和银染获得的电泳谱使菌株能够被评为粗糙、半粗糙或光滑。
We describe a new method for lipopolysaccharide (LPS) preparation by water extraction at 100-degrees-C and subsequent digestion with proteinase K. The crude LPS could be reliably used for immunoblotting since it retained a high level of antigenicity, and was free of SDS and proteinase K, both of which can cause problems. Two monoclonal antibodies which failed to react with LPS prepared by two conventional methods reacted well with our preparation. We used the new method to prepare LPS from 44 strains of bacteria formerly classified as Bacteroides, some of which have been reclassified as Porphyromonas or Prevotella. In general, yields were good, and electrophoretic profiles obtained with SDS-PAGE and silver staining enabled strains to be rated rough, semi-rough, or smooth.