蛋白组学

PROTOCOLTo 1.5 mL eppendorf tubes add:200 µg of protein extract (see Western blot protocol for protein sample preps)q.s. to 300 µL with RIPA (with protease and phosphatase inhibitors).Primary antibody ...

General Preparation1. Prepare buffers and have them cold. Make 50X stock of Buffer G and dilute as needed from frozen aliquots of 50X Buffer G.1X Extraction Buffer without detergents: 1 M Tris-Cl pH 7 ...

co-IP assays can be performed between endogenous proteins or transiently or stably expressed exogenous - usually tagged - proteins. The advantage to using endogenous proteins is the avoidance of prote ...

Western Blot Workflow Diagramback to topProtocolStandard vs. Rapid Immunodetection ProceduresThere are two types of protocols for immunodetection: Standard and rapid. Standard vs. Rapid Immunodetectio ...

1) After ECL development wash membrane once for 10min with PBST.2) Incubate the membrane in stripping buffer (see below) in a heat-sealed plastic bag for 30min at 50oC with occasional mixing. For low- ...

Introduction After proteins have been separated by electrophoresis individual protein bands can often be identified by using an antibody that is specific to that protein. However to be accessible ...

INTRODUCTION In many cases the cleavage can be performed using the free intact fusion or in same cases with the fusion protein bound to a matrix. The amount of thrombin temperature and length of incub ...

INTRODUCTION In many cases the cleavage can be performed using the free intact fusion or in same cases with the fusion protein bound to a matrix. The amount of factor Xa temperature and length of incu ...

Step 1: Transform appropriate DNA plasmid into BL21(DE3) E. coli cells. These cells must be competent. (Protocol for how to make competent cells.) a) Take competent cell stock aliquot (about 100 mL) o ...

MaterialsRIPA buffer (RIPA buffer enables the extraction of cytoplasmic membrane and nuclear proteins and is compatible with many applications including reporter assays protein assays immunoassays and ...

OverviewThis is a protocol for preparing whole cell lysate from tissue for western blot analysis. MaterialsRIPA Bufferper 100ml: (final concentration) 5ml of 1M Hepes pH 7.6 (50 mM) 200µL of 0.5M EDTA ...

Cell Lysis Buffer5mL 0.1M Tris HCl pH 8 (10mM)0.44g NaCl (150mM)0.02g EDTA (1mM)0.5mL nonidet P40 (1% w/v)0.05g SDS (0.1% w/v)Make up to 50mL with MQH2O and filter sterilise.20x Complete mini Protease ...

Cell Lysis Buffer5mL 0.1M Tris HCl pH 8 (10mM)0.44g NaCl (150mM)0.02g EDTA (1mM)0.5mL nonidet P40 (1% w/v)0.05g SDS (0.1% w/v)Make up to 50mL with MQH2O and filter sterilise.Store at 4 degrees C.This ...

General Information Phenol/chloroform extraction is an easy way to remove proteins from your nucleic acid samples and can be carried out in a manner that is very close to quantitative. Nucleic acids r ...

This procedure is suitable for recovering proteins from most aqueous solvents and from SDS containing buffers. It is not recommended for proteins dissolved in urea or guanidine or for peptides.Protoco ...

WB过程中常见的问题及可能原因分析问题可能原因验证或解决办法背景高膜没有完全均匀湿透使用100% 甲醇浸透膜5-10min洗膜不充分增加洗液体积和洗涤次数阻断不充分增加封闭液孵育时间，或者提高温度。选择合适的封闭试剂（脱脂奶粉，BSA，酪蛋白等）二抗浓度过高降低二抗浓度检测过程中膜干燥保证充分的反应液，避免出现干膜现象曝光过度缩短曝光时间抗体与阻断蛋白有交叉反应检测抗体与阻断蛋白的交叉反应性，选择 ...

操作步骤：（实验前请先阅读注意事项）提示：第一次使用前请先在漂洗液RW瓶和70%乙醇瓶中加入指定量无水乙醇!操作前在裂解液RLT中加入β-巯基乙醇至终浓度1%，如1 ml RLT 中加入10μl β-巯基乙醇。此裂解液最好现用现配。配好的RLT 4℃可放置一个月。1.组织培养细胞a.收集b.13，000rpm离心10秒（或者300g离心5分钟），使细胞沉淀下来。完全吸弃上清，留下细 ...

DNA提取次柱漂洗确保高纯度，典型的产量200µl全血可提取出3-6µg，OD260/OD280典型的比值达1.7～1.9，长度可达30 kb -50kb，可直接用于PCR、Southern-blot和各种酶切反应。从十几个配方中优选出的红细胞裂解液配方，裂解快速完全，客户可根据需要选择购买。典型的产量200µl全血可提取出3-6µg 基因组DNA。洗脱液EB不含有螯合剂EDTA，不影响下游酶切、 ...

DNA提取次柱漂洗确保高纯度，典型的产量200µl全血可提取出3-6µg，OD260/OD280典型的比值达1.7～1.9，长度可达30 kb -50kb，可直接用于PCR、Southern-blot和各种酶切反应。从十几个配方中优选出的红细胞裂解液配方，裂解快速完全，客户可根据需要选择购买。典型的产量200µl全血可提取出3-6µg 基因组DNA。洗脱液EB不含有螯合剂EDTA，不影响下游酶切、 ...

使用ProteOn XPR36蛋白相互作用阵列系统分析抗体-膜结合蛋白以及蛋白-脂类物质间的相互作用介绍：研究细胞膜、膜结合蛋白和抗体间的相互作用以及这些作用的互相影响是药物研发中非常值得关注的事情。膜蛋白和膜相关蛋白以及多肽类物质在很多生物过程中的地位都非常重要，同时还参与到一些诸如癌症等疾病的关键信号通路中。因此，膜蛋白正迅速成为下一代药物靶点分子大类。ProteOn XPR36蛋白相互作用阵 ...