目的建立血清蛋白质组学研究中高丰度蛋白的去除方法并评价其去除效果。方法应用Sigma公司2006年最新推出的血清白蛋白和IgG去除试剂盒(ProteoPrep Immunoaffinity Albumin and IgG Depletion Kit)去除血清中高丰度的白蛋白和IgG等,再...目的建立血清蛋白质组学研究中高丰度蛋白的去除方法并评价其去除效果。方法应用Sigma公司2006年最新推出的血清白蛋白和IgG去除试剂盒(ProteoPrep Immunoaffinity Albumin and IgG Depletion Kit)去除血清中高丰度的白蛋白和IgG等,再用双向凝胶电泳(2-DE)分析去除的效果。结果本实验建立的血清中高丰度蛋白的去除方法,约95%的白蛋白和IgG能够被去除,2-DE图谱中蛋白质点更加清楚,部分低丰度蛋白得以显现。结论成功建立血清中高丰度蛋白的去除方法,可为今后的血清蛋白质组学研究奠定基础。展开更多
The aim of this study was to optimize the conditions for the extraction of low-abundance proteins（LAPs） and the removal of abundant proteins（APs; β-conglycinin and glycinin） from soybean meal.Single factor and or...The aim of this study was to optimize the conditions for the extraction of low-abundance proteins（LAPs） and the removal of abundant proteins（APs; β-conglycinin and glycinin） from soybean meal.Single factor and orthogonal experiments were designed to determine the effects of four factors（isopropanol concentration, total extraction time, ultrasonic power, and ultrasonic time） on protein concentration in isopropanol extracts.Proteins in the isopropanol supernatant and the cold acetone precipitate of isopropanol were identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis（SDS-PAGE） and matrix-assisted laser desorption/ionization-time of flight mass spectrometry（MALDI-TOF-MS）.The results showed that the optimal conditions were 50% isopropanol, ultrasonic pretreatment for 15 min at 350 W, and a total extraction time of 1 h.Under these conditions, the protein concentration in the isopropanol extracts reached 0.8081 g/L.Many LAPs were detected, including β-amylase, soybean agglutinin, soybean trypsin inhibitor, fumarylacetoacetase-like, phospholipase D alpha 1-like, oleosin, and even some unknown soybean proteins.The soybean APs（β-conglycinin and glycinin） were not found.The method may be useful for discovering new soybean proteins and extracting enough LAPs of soybean to allow further studies of their physiological effects on animals without the influence of APs.展开更多
目的建立一种去除脑脊液高丰度蛋白的方法并进行双向电泳分析评价。方法采用超滤法浓缩脑脊液样品至适当体积,用商业化血清白蛋白和免疫球蛋白G(IgG)去除试剂盒处理,得到含低丰度蛋白的脑脊液,再用丙酮沉淀法除盐并进行二维凝胶电泳(2-...目的建立一种去除脑脊液高丰度蛋白的方法并进行双向电泳分析评价。方法采用超滤法浓缩脑脊液样品至适当体积,用商业化血清白蛋白和免疫球蛋白G(IgG)去除试剂盒处理,得到含低丰度蛋白的脑脊液,再用丙酮沉淀法除盐并进行二维凝胶电泳(2-DE)分析和2-DE Western blot鉴定。结果此法可有效去除脑脊液中的白蛋白和IgG,2-DE图谱分析显示,去除高丰度蛋白后脑脊液低丰度蛋白富集明显,与2-DE Western blot结果对比,四连接素的三个不同片段去除高丰度蛋白后可全部显现。结论本研究建立的脑脊液高丰度蛋白去除方法简便易行,有利于双向电泳中低丰度蛋白的检出。展开更多
文摘目的建立血清蛋白质组学研究中高丰度蛋白的去除方法并评价其去除效果。方法应用Sigma公司2006年最新推出的血清白蛋白和IgG去除试剂盒(ProteoPrep Immunoaffinity Albumin and IgG Depletion Kit)去除血清中高丰度的白蛋白和IgG等,再用双向凝胶电泳(2-DE)分析去除的效果。结果本实验建立的血清中高丰度蛋白的去除方法,约95%的白蛋白和IgG能够被去除,2-DE图谱中蛋白质点更加清楚,部分低丰度蛋白得以显现。结论成功建立血清中高丰度蛋白的去除方法,可为今后的血清蛋白质组学研究奠定基础。
基金Project supported by the China Agriculture Research System(No.CARS-36)the National Natural Science Foundation of China(No.31572430)
文摘The aim of this study was to optimize the conditions for the extraction of low-abundance proteins（LAPs） and the removal of abundant proteins（APs; β-conglycinin and glycinin） from soybean meal.Single factor and orthogonal experiments were designed to determine the effects of four factors（isopropanol concentration, total extraction time, ultrasonic power, and ultrasonic time） on protein concentration in isopropanol extracts.Proteins in the isopropanol supernatant and the cold acetone precipitate of isopropanol were identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis（SDS-PAGE） and matrix-assisted laser desorption/ionization-time of flight mass spectrometry（MALDI-TOF-MS）.The results showed that the optimal conditions were 50% isopropanol, ultrasonic pretreatment for 15 min at 350 W, and a total extraction time of 1 h.Under these conditions, the protein concentration in the isopropanol extracts reached 0.8081 g/L.Many LAPs were detected, including β-amylase, soybean agglutinin, soybean trypsin inhibitor, fumarylacetoacetase-like, phospholipase D alpha 1-like, oleosin, and even some unknown soybean proteins.The soybean APs（β-conglycinin and glycinin） were not found.The method may be useful for discovering new soybean proteins and extracting enough LAPs of soybean to allow further studies of their physiological effects on animals without the influence of APs.
文摘目的建立一种去除脑脊液高丰度蛋白的方法并进行双向电泳分析评价。方法采用超滤法浓缩脑脊液样品至适当体积,用商业化血清白蛋白和免疫球蛋白G(IgG)去除试剂盒处理,得到含低丰度蛋白的脑脊液,再用丙酮沉淀法除盐并进行二维凝胶电泳(2-DE)分析和2-DE Western blot鉴定。结果此法可有效去除脑脊液中的白蛋白和IgG,2-DE图谱分析显示,去除高丰度蛋白后脑脊液低丰度蛋白富集明显,与2-DE Western blot结果对比,四连接素的三个不同片段去除高丰度蛋白后可全部显现。结论本研究建立的脑脊液高丰度蛋白去除方法简便易行,有利于双向电泳中低丰度蛋白的检出。
