Nativechromatinimmunoprecipitationprotocol
实验概要The method is a native chromatin immunoprecipitation protocol.主要试剂1. 10 x TBS 0.1 M Tris-HCl (pH 7.5) 1.5 M NaCl 30 mM CaCl2 20 mM MgCl2 50 mM Na butyrate (pH 8.0)2. Digestion buffer 0.32 M sucrose 50 mM Tris-HCl (pH 7.5) 4 mM MgCl2 1 mM CaCl2 0.1 mM PMSF 5 mM Na butyrate3. Lysis buffer 1.0 mM Tris-HCl (pH7.4) 0.2 mM Na2EDTA 0.2 mM PMSF 5 mM Na butyrate4. Incubation buffer 50 mM NaCl 20 mM Tris-HCL (pH 7.5) 20 mM......阅读全文
ChIPChip－E.－coli
AbstractChIP-Chip stands for Chromatin Immunoprecipitation and chip in the sense of DNA microarray. It is a technique to determine the genome-wide bin
GFPTrap如何设计成功的IP实验？
借助Chromotek公司GFP-Trap如何设计成功的IP实验？How to plan an immunoprecipitation of your GFP-fusion protein when using the ChromoTek GFP-Trap®PreambleThis document
Fragment－Complementation－and－Coimmunoprecipitation－Assays－for－...
Plant disease resistance (R) proteins confer protection against specific pathogens or pathogen isolates. R proteins function by recognizing pathog
Native－gel－electrophoresis（非变性电泳）
Native gel electrophoresis Under native PAGE conditions, polypeptides retain their higher-order structure and often retain enzymatic activity and inte
Chromatin－Remodeling－by－hSWI/SNF－ATPdependent－Complexes
The eukaryotic genome is packaged by histone and nonhistone proteins to form chromatin. The assembly of nucleosomesas well as compaction of nucleosoma
ELISPOT－Protocol
实验概要The Elispot (Enzyme Linked Immuno-Spot) assay provides an effective method of measuring the antibody or cytokine production of immune cells on t
Immunoblot－Protocol
This method was successful in our lab using prostate tissue and for our specific objectives. Investigators must be aware that they will need to tailor
ELISPOT－protocol
实验概要The procedure below is a general guideline procedure for ELISPOT. Abcam ELISPOT kits have been designed for detection of various cytokines and g
RLGS－protocol
A. Preparation of DNA SolutionIn the case of rice, for example This method may be appllicable for many grass species and some other plants.
ELISA－protocol
ELISA protocol：1.取5－10ul BMMY表达上清用0.05M NaHCO3稀释到100ul铺ELISA板，37度或室温振荡大于1小时。注意一定要做一个GS115空菌株表达上清作为阴性对照，最好还找一个带有histag的蛋白作为阳性对照。2.TPBS洗板3次，方法：倒掉铺板液，倒置于
NAi－protocol
siRNA protocolsOur current strategy with siRNA is to synthesis relatively small amounts enzymatically and use these to test for efficiency by western
PCR－protocol
PCR reactionProtocol for 50µl reaction - adjust amounts if necessary, for a 20µl reaction use the same volumes of primer and dNTP-mix, but adjust the
RNAi－protocol
siRNA protocolsOur current strategy with siRNA is to synthesis relatively small amounts enzymatically and use these to test for efficiency by western
ELISPOT－Protocol
实验概要The Elispot (Enzyme Linked Immuno-Spot) assay provides an effective method of measuring the antibody or cytokine production of immune cells on t
蛋白质磷酸化
Tyrosine Kinase Assay Using Synthetic Peptides (T. Miller)Small synthetic peptide substrates are especially well suited for applications such as assay
(NativePAGE)应该注意哪些问题
1. 非变性聚丙烯酰胺凝胶电泳的过程中，蛋白质的迁移率不仅和蛋白质的等电点有关，还和蛋白质的分子量以及分子形状有关，其中蛋白质的等电点是最重要 的影响因子，要根据蛋白质的等电点来选择对应的电泳缓冲系统； 2. 非变性聚丙烯酰胺凝胶电泳的过程中，要注意电压过高引起发热而导致蛋白质变性，所以最好在电泳槽
酵母染色体沉淀分析方法
ABSTRACTThis protocol describes a method for the detection of proteins bound to specific regions of chromatin in yeast. There are many variations of t
胞外基质
ECM Cell Attachment Assay (LTI)Cell Adherence Inhibition Assay (LTI)General protocol--Either monoclonal antibody or RGD peptide is added along with th
免疫沉淀（Immunoprecipitation）实验材料和方法
免疫沉淀是利用特异性抗体识别并分离抗原的方法。材料：上样buffer的配方(用前现配):2ul 1.25M Tris-HCL , pH 6.835ul distilled water2.5ul 2-mercaptoethanol12.5ul 10%SDS10ul 80% glycerol2ul br
免疫沉淀（Immunoprecipitation,－IP）实验方法
基本实验步骤 （1）收获细胞，加入适量细胞IP裂解缓冲液（含蛋白酶抑制剂），冰上或者4℃裂解30min, 12,000g离心30 min后取上清； （2） 取少量裂解液以备Western blot分析，剩余裂解液将1μg相应的抗体和10-50 μl protein A/G-beads加入到细胞裂解
多名学者推荐全基因组分析转录因子功能新技术
来自北卡罗来纳大学教堂山分校，NIH等处的研究人员利用一种新型方法，分析了酵母转录因子Rap1在整个基因组中的结合动态，从而可以更好研究这一转录因子的功能，这一方法将有助于科学家们实时分析转录情况，相关成果公布在Nature杂志上。 对于这一成果，来自法国国家科学研究中心的François
Migration－Assay－Protocol
Materials to be prepared beforehand:1) FBS free medium2) 10% FBS medium3) Cell migration filter insert ( Transwell®, 12mm Diameter, 12 μm Pore Size.)P
Protocol－for－Trichl...
实验概要The efficiency of nucleotide incorporation in DNA/RNA polymerization reactions (e.g. transcription, reverse transcription, and DNA replication)
cDNA/AFLP－Protocol
Preparation of Para-magnetic beads from Promega cat#Z5482:a) suspend magnetic particles in bottle - transfer 200 ul (200 ug) of beads per sampleof RNA
Urea－Lysis－Protocol
Urea lysis buffer 9M Urea, 2.5mM EDTA, 2.5mM EGTA, 1% DTE, 4% CHAPS make 10ml and aliquot 10x1ml, freeze at -70°C Lysate prepara
Protocol－for－Cell－Fusion
Healthy Sp2/0 cells should be rapidly growing by this time. Sp 2/0 cells should be started about two weeks before the cell fusion. Every two days, the
Dot－Blot－Protocol
a. Label nitrocellulose membrane (using a pencil) to identify protein elution fractions.b . Pipette 2μl from each fraction onto the membrane, allow th
TAIL－PCR－Protocol
TAIL is a series of reactions that are intended to map where a T-DNA (transfer DNA) has inserted within the genome. The main components of the 3 react
Intracellular－Staining－Protocol
1. Fix cells- Add16% formaldehyde directly into culture medium to obtain a final concentration of 1.5% formaldehyde.2. Incubate in fixative for 10 min
Cell－Extraction－Protocol
实验概要Primary tissues are valuable tools for the study of intracellular and extracellular markers which characterize disease states. We have developed