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=== BCA定量蛋白 === *'''400000细胞密度接种于6孔细胞培养板,分别于2ml RPMI完全培养基(PBS、LMWH、AST、LANPs(ug/ml))中培育12h''' *'''收集细胞,用细胞核/细胞质蛋白提取试剂盒分别提取各组蛋白质样品(用RIPA裂解缓冲液裂解细胞20 mg--100~200 ul),(4℃10000-14000离心10-15 min),用BCA蛋白浓度测定试剂盒对蛋白质进行浓度定量''' *'''蛋白样品与SDS-PAGE上样缓冲液(5×)按体积比4:1均匀混合,沸水浴10min,冷却后,将各组蛋白用缓冲液(1×)稀释至同一浓度备用(-80℃保存)''' *'''400000细胞密度接种于6孔细胞培养板,分别于2ml RPMI完全培养基(PBS、LMWH、AST、LANPs(ug/ml))中培育12h''' *'''收集细胞,用细胞核/细胞质蛋白提取试剂盒分别提取各组蛋白质样品(用RIPA裂解缓冲液裂解细胞20 mg--100~200 ul),(4℃10000-14000离心10-15 min),用BCA蛋白浓度测定试剂盒对蛋白质进行浓度定量''' *'''蛋白样品与SDS-PAGE上样缓冲液(5×)按体积比4:1均匀混合,沸水浴10min,冷却后,将各组蛋白用缓冲液(1×)稀释至同一浓度备用(-80℃保存)''' <br> <div style="color: red !important> *'''''BCA:A液 B液 蛋白标准样品(蛋白浓度0125-2 mg/ml)''''' *'''''测定:a.每孔加入10 ul样品(96孔板) b.加入200 ul工作液振摇(30 S) c.37℃孵育0.5h(时间越久,温度越高,测量值越高) d.562nm酶标仪测量 e.标准曲线 f.恒温加热器100℃ 5-10 min ,分装保存(-40℃)''''' <div style="color: black !important; font-family: 'Times New Roman', SimSun, serif !important, line-height:1.5;">
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