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Western Blotting with Biotinylated Antibodies SAMPLE PREPARATION For Protein Concentration Determination of Cell Culture 1.Decant medium from 10cm dish of adherent cells and rinse plate rapidly with p ...

SAMPLE PREPARATION For Protein Concentration Determination of Cell Culture 1.Decant medium from 10cm dish of adherent cells and rinse plate rapidly with phosphate-buffered saline (PBS). 2.Aspirate ex ...

Sample Preparation For Protein Concentration Determination of Cell Culture 1.Decant medium from 10cm dish of adherent cells and rinse plate rapidly with phosphate-buffered saline (PBS). 2.Aspirate e ...

Running Protein Gels Solutions 10X Running Buffer (0.25 M Tris 1.92 M glycine 1% SDS) 121 g Tris 577 g glycine 40 g SDS ddh20 to 4 L (check pH at 1:10 dilution (pH=~8.8)). 5X Sample Buffer (0.3125 M T ...

Studying Protein Interactions by Far-Western Blotting Far-Western blotting was originally developed to screen protein expression libraries with 32P-labeled glutathione S-transferase (GST)-fusion prot ...

1. Run samples out on a gel. For bacterially expressed proteins generally 5 μl is plenty (1ml cell cμlture; cells resuspeded in 50 μl loading buffer). Run the gels (BioRad mini gels) at 195 V ...

1.Remove the stacking gel from the gel. Measure the gel and record its size. Do not pre-soak the gel. Do not cut a corner off the gel as this may allow a short circuit and inefficient transfer. Blot t ...

1. 30%储备胶溶液： 丙烯酰胺（Acr） 29.0g　　　　　　　　　　　亚甲双丙烯酰胺（Bis） ...

Storage of lyophilized antibodies All our products are shipped lyophilized (freeze-dried). Unlabeled antibodies are stable without loss of quality at ambient temperatures for several weeks or even a ...

0 128); font-weight: bold;"Western免疫印迹（Western Blot）是将蛋白质转移到膜上，然后利用抗体进行检测。对已知表达蛋白，可用相应抗体作为一抗进行检测，对新基因的表达产物，可通过融合部分的抗体检测。 0 128);"原理与Southern或Northern杂交方法类似，但Western Blot采用的是聚丙烯酰胺凝胶电泳，被检测物是蛋白质，“ ...

Western- or immunoblotting is a commonly employed technique for the detection of protein antigens in complex mixtures. Samples are first separated by SDS-polyacrylamide gel electrophoresis. The separa ...

1. Perform SDS-polyacrylamide gel electrophoresis (SDS-PAGE) on a cell sample prepared using modified and transfer the proteins to a nitrocellulose membrane. 2. Wash the nitrocellulose twice with dis ...

我是做临床的，07年毕业。 06年完成老板的RNA干扰体内体外课题，实验花了7个月，大家都说我神速，我也觉得也是快了些，呵呵，看到其他同学忙得没结果，我觉得我是幸运的。自己还得了优秀毕业论文。在感激老天眷顾、高兴之余我想和大家分享自己做实验的体会： 一、在选题方面要注意：根据自己实验条件，来选择实验内容及检测指标，这一点很重要，好多战友在选题上难度很大，自己操作起来很难，把试剂买来了，做了太难，放 ...

Use alternate buffer system (such as CAPS buffer pH 10.5) with a higher pH. Assemble transfer stack with additional membrane soaked in cathode buffer on the cathode side of gel. This will captur ...

Western Blot详解(原理、分类、试剂、步骤及问题解答)

0 128);"REAGENTSECL Western blotting kit (Amersham Life Science; cat# RPN2108): contains second antibodies for both mouse and rabbit substrate and milk blocker (the milk blocker is not normally used ...

Solutions: Buffer A (Hypotonic Buffer): 1L 10 mM HEPES pH 7.9 10 ml 1M HEPESpH 7.9 1.5 mM MgCl2 1.5 ml 1M MgCl2 10 mM KCl3.33 ml 3M KCl 0.5 mM DTT 500 µl 1M DTT 0.2 mM PMSF 1 ml 0.2 M PMSF Buffe ...

（一）双缩脲测定法 1．原理 蛋白质中的肽键有双缩脲反应，在碱性溶液中与二价铜离子形成蓝紫色的络合物，在一定的范围内，颜色的深浅与蛋白质的含量成正比。此法特异性强，游离的氨基酸、小肽和核酸均不产生这种反应，但此法不够敏感，仅能测出毫克水平。 2．试剂配制 硫酸铜（CuSO4·5H2O） 1.50g 酒石酸钾钠 5.00g H2O 500.0ml 10%氢氧化钠（不含硫酸钠）300m ...

Grow a 2 ml culture24 hr.at 30℃ in selective media When culture is readyuse it to inoculate about 55 ml (50 ml plus 5 for O.D.600 readings)of selective media.Grow to mid-logarithmic phase (O.D.600 is ...

随着分子生物学的发展，越来越多的科研人员熟练掌握了分子生物学的各种试验技术，并研制成套试剂盒，使基因克隆表达变得越来越容易。但分子生物学的上游工作往往并非是最终目的，分子克隆与表达的关键是要拿到纯的表达产物，以研究其生物学作用，或者大量生产出可用于疾病治疗的生物制品。相对与上游工作来说，分子克隆的下游工作显得更难，蛋白纯化工作非常复杂，除了要保证纯度外，蛋白产品还必须保持其生物学活性。纯化工艺必须 ...