Incorrect storage of antibodies or ECL western blotting detection reagents may result in a loss of signal. Insufficient protein may have been loaded on the gel. Depending on the location of the target protein, membrane or nuclear preparations may be required (instead of whole cell lysates). Film...
4、 封闭:电转后膜用1XTBS漂洗1次,置于5%脱脂奶粉中(1XTBS+1000:1 TWEEN20),封闭过夜,或者37℃,30 min ~90 min。 5、 一抗:抗体(抗体稀释比例根据自身实验具体确定,推荐比例:抗血清常用1:100~1:5000 5%牛奶,培养液上清1:2~1:100,腹水1:200~1:10000 5%牛奶,),震荡孵育1或2小时。(一抗可回收冷冻...
Western Blotting Protocol 蛋白印迹实验步骤 蛋白印迹实验 (WB) 广泛用于分析细胞或组织提取物中的特异性蛋白表达。我们理解蛋白印迹实验非常耗时,且对于您的研究进展至关重要。为此,我们在提供优化的实验步骤时,还提供参考信息和技术支持,以便让您在蛋白印迹实验中取得成功。
Western blotting Protocol(试剂配方和实验操作) 一、 试剂配方: 1X TBS: Tris-base 12.11g Nacl 8.775g 用HCl 调pH7.4,用纯水稀释至1L 5X Transfer buffer: Tris-base 15.1g Glycine 72.0g To 1 L 10X碱性磷酸酶缓 一、 试剂配方: 1X TBS: Tris-base 12.11g Nacl 8.775g 用HCl 调pH7.4,用纯水稀释至...
Western blotting (WB) is widely used to analyze specific protein expression in cell or tissue extracts. At Cell Signaling Technology (CST) we understand that western blotting experiments are time consuming and that their success has a critical impact on your research progress. For that reason, we...
Western Blotting Protocol-中山大学(参考)Western Bloting(张雪雁) 操作步骤: 1.做胶:下层胶7.5ml一块,以水或乙醇隔离空气。待下层胶凝固后做上层胶,每块胶配置3ml,灌胶、插梳子,赶走气泡。 2.蛋白变性:在胶凝固过程中,变性蛋白质,蛋白质的上样量按事先所测弄浓度计算准确,分装到EP管或,按5ul/100ul样品的...
Western blotting protocol By Wei-Zhang Wang-070610 A. 溶液和试剂 注意:溶液的配制最好用 Milli-Q 或同等纯度的水!所有下面所用容器使用前都必须洗干净,并用双 蒸水润洗,最后晾干。 A1. 抑制剂: 蛋白酶抑制(35mg/ml PMSF):用无水乙醇稀释为 35mg/ml,室温下避光保存,可使 用 3-4 个月。(MW:172.4...
WesternblottingprotocolWesternblottingprotocolWesternblottingprotocol ByWei-ZhangWang-070610ByWei-ZhangWang-070610ByWei-ZhangWang-070610ByWei-ZhangWang-070610 A.A.A.A. Milli-QMilli-QMilli-QMilli-Q!!! A1.A1.A1.抑制剂:抑制剂:抑制剂: 蛋白酶抑制(蛋白酶抑制(蛋白酶抑制(35mg/mlPMSF35mg/mlPMSF35...
Because amino acid sequences vary from protein to protein, western blotting analysis can be used to identify and quantify a single protein in a lysate that contains thousands of different proteins. First, proteins are separated from each other based on their size by SDS-PAGE gel electrophoresis....
westernblottingprotocol Western-blotting 1.从植物材料提取蛋白质 取一些植物材料用液氮速冻后放入研钵研磨成粉末,加入5%SDS(每100mg 植物材料加50 μl 10%SDS和50ul蒸馏水),沸水浴3min,13000rpm离心3min,吸取上清后加入等体积2×SDS loading buffer(注意加入DTT),沸水浴3min。 换用extraction buffer (150 mM Tris...