CoIPProtocol1
一 原理：IP是利用抗原蛋白质和抗体的特异性结合以及细菌蛋白质的“prorein A"特异性地结合到免疫球蛋白的FC片段的现象活用开发出来的方法。目前多用精制的prorein A预先结合固化在argarose的beads上，使之与含有抗原的溶液及抗体反应后，beads上的prorein A就能吸附抗原达到精制的目的。实验最需要注意点就是抗体的性质。抗体不同和抗原结合能力也不同，免染能结合未必能用在IP反应。建议仔细检查抗体的说明书。特别是多抗的特异性是问题。其次，要注意溶解抗原的缓冲液的性质。多数的抗原是细胞构成的蛋白，特别是骨架蛋白，缓冲液必须要使其溶解。为此，必须使用含有强界面活性剂的缓冲液，尽管它有可能影响一部分抗原抗体的结合。另一面，如用弱界面活性剂溶解细胞，就不能充分溶解细胞蛋白。即便溶解也产生与其它的蛋白结合的结果，抗原决定族被封闭，影响与抗体的结合，即使IP成功，也是很多蛋白与抗体共沉的悲惨结果。再次，为......阅读全文
RNA－Isolation－Protocol
Stabilize RNAStart with 15 ml E. coli Culture containing 7.5* 109 cells (OD600= 0.2 Dilute cells or scale up)Pipet 30 ml of RNAProtect Bacteria Reagen
Western－Blot－Protocol
一、提取抗原蛋白将提取RNA途中留存的样品，加入150μl 100%酒精充分混匀，静置5min(RT), 2000×g , 4℃离心5min, 吸取上清至新管中, 加入750μl异丙醇, 混匀, 静置10min(RT), 12000×g, 4℃离心10min, 弃上清, 加入1ml 0.3mol/L
Microarray－Hybridization－Protocol
Introduction:One microarray set consists of 7 nylon membranes with 2.5 x 7.5 cm dimension. 2304 genes were spotted onto nylon membranes (Schleicher an
Histone－blotting－protocol
实验概要 Western blot detection of histone proteins. 实验步骤 The following protocol refers to the western blot detection of histone proteins derived from p
Bacteria－Culture－Protocol
Bacteria Culture ProtocolBy 徐晓政1、TBS Medium Preparation:Prepare 1L of TBS medium contains:Tryptone 12gYeast extract 24gNaCl 5gSodium Succinate 5gGlyce
IP防水防尘测试方法
IP6X将样品于接头组装成一体放入防尘箱中，测试中应用滑石粉，测试时间：8 小时没有灰尘侵入IPX8将样品与接头组装成一体装入试验工装内，从外接气插口向实验工装内注入清水，水面高于试件顶部至少20mm，水面上方留有一定的空间，在进气插头上插入进气管，另在工装漏水观察胶管插头接100mm 短管，从进气
大鼠IP3(IP3)ELISA试剂盒使用说明
原理本实验采用双抗体夹心 ABC-ELISA法。用抗大鼠 IP-3(interferon-inducible protein 3) 单抗包被于酶标板上，标准品和样品中的 IP-3与单抗结合，加入生物素化的抗大鼠IP-3，形成免疫复合物连接在板上，辣根过氧化物酶标记的Streptavidin与
大鼠IP10(IP10)ELISA试剂盒使用说明
原理本实验采用双抗体夹心 ABC-ELISA法。用抗大鼠 IP-10(interferon-inducible protein 10) 单抗包被于酶标板上，标准品和样品中的 IP-10与单抗结合，加入生物素化的抗大鼠IP-10，形成免疫复合物连接在板上，辣根过氧化物酶标记的Streptavi
Streamlined－DNA－Extraction－Protocol
This method is derived from a procedure developed by Toby Bradshaw and the Poplar Molecular Genetics Cooperative. We have tested the procedure wit
Xenograft－Tumor－Assay－Protocol
1) Determine the number of cells for injection (ie 5´106 ) to determine the number of plates thatwill require trypsinizing (usually a 100% confluent p
ELISA－Protocol－(General－Guidelines)
实验概要Sandwich enzyme-linked immunosorbent assays (ELISAs) involve attachment of a capture antibody to a solid phase support. Samples containing known
Native－chromatin－immunoprecipitation－protocol
实验概要Native chromatin immunoprecipitation to query specific chromatin states of individual genes. 主要试剂10 x TBS 0.1 M Tris-HCl (pH 7.5) 1.5 M NaCl 30 mM
Simplified－Arabidopsis－Transformation－Protocol
(Brief version for those who are familiar with the method)Steve Clough and Andrew Bent, University of Illinois at Urbana-Champaign.Our present proto
Immunohistochemistry－Protocol－for－Frozen－Sections
实验概要The following is a general procedure guide for preparation and staining of acetone-fixed frozen tissues using a purified, unconjugated primary
The－protocol－for－LIC－by－Exonuclease－III
The protocol for LIC by Exonuclease III梁耀极1. Design the primers with 15-bp overlap;2. Digest the vector by proper restriction enzyme;For getting high
Intracellular－Cytokine－Staining－Protocol
实验概要A modification of the basic immunofluorescence staining and flow cytometric analysis protocol can be used for the simultaneous analysis of surf
Blocking－with－immunizing－peptide－protocol
实验概要The method provides a blocking with immunizing peptide protocol.实验原理Non-specific binding of an antibody to proteins other than the antigen can s
Tissue－preparation－protocol－for－ChIP
实验概要This protocol describes how chromatin is prepared from tissue, which can subsequently be used for chromatin immunoprecipitation (ChIP). It is re
RTPCR－PROTOCOL
RT-PCR PROTOCOL材料与方法………………………………………………………… 1．材料 ………………………………………………………1.1 供试用组织(细胞)…………………………………1.2 主要仪器设备………………………………………1.3 主要试剂……………………………………………1.
Arabidopsis－RNA－extraction－protocol
1-2 g fresh material, freezer-dried, ground with 0.2g sand (if necessary), and then homogenized with 10ml RNA extraction buffer (see below). Spin
Immunohistochemistry－Protocol－for－Frozen－Sections
实验概要The following is a general procedure guide for preparation and staining of acetone-fixed frozen tissues using a purified, unconjugated primary
Protocol－for－Construction－of－BAC－Libraries
Protocol for Construction of BAC Libraries The bacterial artificial chromosome cloning (BAC) system is emerging as the system of choice for const
Cajal－Body－Isolation－Protocol
Buffers and solutions(All solutions are supplemented with Complete Protease inhibitor tablet (Roche, Cat no: 1-873-580) at the final concentration of
Protocol－for－Aortic－Ring－Assay
ProceduresCover a 48-well plate with Matrigel (100μl/well) of and incubate for 30 min at 37°C, 5% CO2.Sacrifice the1-2 month old mice/rats (WT/mutant
Immunofluorescence－/－Confocal－Microscopy－Protocol
实验概要Immunofluorescence is a technique used for light microscopy with a fluorescence microscope and is used primarily on biological samples. This tec
ELISA－Protocol－(General－Guidelines)
实验概要Sandwich enzyme-linked immunosorbent assays (ELISAs) involve attachment of a capture antibody to a solid phase support. Samples containing known
stem－cell－culture－protocol
实验概要stem cell culture protocol主要试剂cell culture supplies and reagentssEnvironment: cell culture requires a sterile environment, so it needs a separat
Immunofluorescence－/－Confocal－Microscopy－Protocol
实验概要Immunofluorescence is a technique used for light microscopy with a fluorescence microscope and is used primarily on biological samples. This tec
Native－chromatin－immunoprecipitation－protocol
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
Simplified－Arabidopsis－Transformation－Protocol
实验概要Our present protocol (Clough and Bent, 1998; modified from Bechtold et al. 1993) is extremely simple. We have found that the MS salts, hormone