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巨噬细胞源性神经营养因子的纯化和鉴定 被引量:4
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作者 李松林 郭畹华 顾熊飞 《实验生物学报》 CSCD 1998年第2期137-145,共9页
应用Sephacryl S-100-HR凝胶层析、高效液相色谱(HPLC)和反相高效液相色谱(R-HPLC)分析,从巨噬细胞条件培养基中纯化了巨噬细胞源性神经营养因子(MΦDNF)。该因子在SDS-聚丙烯酰胺凝胶电泳上呈现一条单一蛋白带,其分子量为60.5kD,等电点... 应用Sephacryl S-100-HR凝胶层析、高效液相色谱(HPLC)和反相高效液相色谱(R-HPLC)分析,从巨噬细胞条件培养基中纯化了巨噬细胞源性神经营养因子(MΦDNF)。该因子在SDS-聚丙烯酰胺凝胶电泳上呈现一条单一蛋白带,其分子量为60.5kD,等电点约5.1。其氨基酸组成含有较高比例的天冬氨酸、谷氨酸、亮氨酸和赖氨酸。纯化的MΦDNF能支持体外培养的小脑皮质神经元的存活,增强其活性及促进神经元突起的生长。其最大效应浓度为500—1000ng/ml。 展开更多
关键词 巨噬细胞源性 神经营养因子 纯化 鉴定
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巨噬细胞源性神经营养因子的初步分离
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作者 李松林 郭畹华 顾熊飞 《解剖科学进展》 CAS 1996年第4期363-369,共7页
本文制备了巨噬细胞条件培养基(MφCM),并应用快速自动比色微量分析法检测MφCM对体外培养的生后7dSD大鼠小脑皮质神经元的作用。结果表明,分子量大于10kD的MφCM对神经元(细胞密度1×106/ml)的作用... 本文制备了巨噬细胞条件培养基(MφCM),并应用快速自动比色微量分析法检测MφCM对体外培养的生后7dSD大鼠小脑皮质神经元的作用。结果表明,分子量大于10kD的MφCM对神经元(细胞密度1×106/ml)的作用,与对照组比较有明显的神经营养活性(F<0.05);用盖玻片培养法表明,该组份有促进神经元突起生长的作用。MφCM经SephacrylS-100-HR凝胶层析和生物活性鉴定,获得了具有神经营养活性的第二峰洗脱液。此洗脱液经SDS-聚丙烯酰胺凝胶电泳分析,证明MφCM中神经营养活性成份的蛋白质其分子量为31kD-68kD之间。 展开更多
关键词 巨噬细胞源性 神经营养因子 MΦCM 小脑皮质 神经元 细胞培养 Sephacreyl s-100-hr凝肢层析
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Isolation and characterization of a new subunit of phycocyanin from Chroomonas placoidea 被引量:7
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作者 Yun Yun Zhang Min Chen Hong Cui 《Chinese Chemical Letters》 SCIE CAS CSCD 2011年第10期1229-1232,共4页
A new phycocyanin(PC) fluorescent subunit namedβ_2(18kDa) was isolated and characterized by both SDS-PAGE and isoelectric focusing(IEF) from a species of cryptophytic alga Chroomonas placoidea.PC was separated ... A new phycocyanin(PC) fluorescent subunit namedβ_2(18kDa) was isolated and characterized by both SDS-PAGE and isoelectric focusing(IEF) from a species of cryptophytic alga Chroomonas placoidea.PC was separated and purified by ammonium sulfate sedimentation followed by two steps of Sephadex G-100 chromatography.After denatured in 4 mol/L urea for 48 h,PC was divided into two fractions by passing through a Sephacryl S-100 chromatography column twice.The blue fraction(S-1) containedβsubunits with a maximal absorbance at 595 nm in visible light region.While the green fraction(S-2) enriched inαsubunits showed a characteristic long wavelength absorbance at 680-700 nm region and exhibited a relatively low molecular weight of 9.4(α_1) and 8.5 kDa(α_2).Fraction S-1 also consisted of two different fluorescent subunits with molecular weight of 20.1 kDa(β_1) and 18 kDa (β_2) and differed from each other on isoelectric points of pH 5.7(ft) and 6.0(ft),respectively.Further investigation of peptide sequence will help a lot in elucidating the new subunit ft that was smaller in size and more neutral than the known ft subunit,and may provide an alternative explanation in structure of cryptophytic phycobiliproteins. 展开更多
关键词 Chroomonas placoidea Phycocyanin subunit sephacryl s-100 chromatography SDs-PAGE IEF
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