Gelfiltration装填的基本原理
Gel filtration的基本原理，现在谈谈我装柱子的过程。我们用的是Amersham 的 XK26/40的空柱子，然后往里面填充Sephacryl S-300 HR 的resin。按道理说，要提高分辨率，柱子应该是越长越细才好，这种柱子虽然不短，但是挺粗的。但是一个好处就是，样品体积可以大一些。 装柱子本身就是一件很tricky的事情，以至与这课的教程上专门列了一小节来解释怎么装柱子。我们这一组人中，我负责装柱子。一开始就是洗resin了，用粗制烧结玻板过滤的漏斗来洗，感觉这方法还挺快的。洗完了，就把resin重悬成50%的slurry，然后就是往柱子里头倒了。但是呢，有一个小窍门，就是必须先封闭柱子的下出口，往柱子里面倒10%体积的buffer。我过去装柱子犯傻，不知道要这么做，结果发现下面的resin 不均匀不说，还有一大堆气泡。所以先加buffer是很必要的。倒slurry的时候还得用玻棒引流，这样可以......阅读全文
InGel－Digestion－of－Proteins－Separated－byPolyacrylamide－Gel－Electrophoresis
1. Excision of protein bands (spots) from polyacrylamide gelsRinse the gloves you use with water to avoid traces of dust in your sample.Rinse the gel
Gel－Electrophoresis－of－DNA
What is Electrophoresis?Electrophoresis is a technique used in the laboratory that results in the separation of charged molecules. In this CyberLab we
RNA－gel－electrophoresis
实验概要RNA gel electrophoresis主要试剂DEPC H2ODEPC 0.1% (v/v)q.s. de-ioinized H2O37ºC x1 hr, or r.t. overnightAutoclave.(NaOAc, EDTA and ethidium bromide sol
RNA－gel－electrophoresis
MaterialsDEPC H2ODEPC 0.1% (v/v)q.s. de-ioinized H2O37ºC x1 hr, or r.t. overnightAutoclave.(NaOAc, EDTA and ethidium bromide solutions should also be
Agarose－Gel－Electrophoresis
实验概要Separating nucleic acid fragments by agarose gel electrophoresis.实验原理 Agarose gel electrophoresis remains the most widely used technique for sep
Agarose－gel－electrophoresis
General ProcedureCast a gelPlace it in gel box in running bufferLoad samplesRun the gelImage the gelCasting Gels0.7% agarose gel with 1kbp ladder in U
Agarose－Gel－Electrophoresis－of－DNA
1) Dissolve 1 g of agarose in 100 ml of 1X TAE or TBE buffer (gives a 1% gel). See note for making LMP agarose gel. 2) Cast the gel with the comb in p
Polyacrylamide－Gel－Electrophoresis－of－Oligonucleotides
1. Pour and polymerize a 20% polyacrylamide gel, no Urea.2. Remove clamps. Rinse with water. Remove comb. Rinse top of gel well.3. Insert comb teeth d
Alkaline－agarose－gel－electrophoresis
Alkaline agarose gel electrophoresis (Sambrook et al., 1989)Alkaline agarose gels can be used to determine the size and quality of first and second st
Gel－Shift－Assay－Systems
ProtocolsDownloadprotocol183kbpdf?Abstract for Gel Shift Assay SystemsThe gel shift, or electrophoretic mobility shift, assay provides a simple and ra
Blue－Native－Gel－Electrophoresis
Blue Native Gel ElectrophoresisStock solutions49.5%T, 3%C Acrylamide 24 g acrylamide, 0.75 g bisacrylamide / 50 ml H2O Store at RT3 x Gel buffer 150 m
SDS－Gel－Electrophoresis－of－Tubulin\MAPs
MaterialsStock Acrylamide: (30%T:0.8%C)30% by weight of acrylamide0.8% by weight of N,N'-bis-methylene acrylamideSeparation Gel (Final Concentrati
High－Resolution－Agarose－Gel－Electrophoresis
实验概要Agarose gel electrophoresis remains the most widely used technique for separating nucleic acid fragments due to its ease of use, non-toxicity, a
Acrylamide－Urea－Gel－(35－ml)
Acrylamide Urea Gel (35 ml)10%15%40/2% acrylamide10 ml13.1 ml10X TBE3.5 ml3.5 mlUrea15 g15 gH2010ml7.0 ml Microwave ~10 seconds and stir until dissolv
Protein－concentration－of－Laemmli－gel－samples
Protein concentration of Laemmli gel samplesTo 10 µl boiled lysate (in Laemmli sample buffer) add 40µl water + 50µl 50% TCA. Ppt. 10 min. on ice. Spin
SSR－GEL－and－Silver－Staining－Protocol
I. EQUIPMENT:DNA sequencing unit (35 x 45 cm) & 2000V power supplyClampsLg. plastic trays (4), about 43 x 50 x 8 cm, and one lidTwo rocking platformsH
Denaturing－Agarose－Gel－Electrophoresis－of－RNA
The overall quality of an RNA preparation may be assessed by electrophoresis on a denaturing agarose gel; this will also give some information about R
Denaturing－Gradient－Gel－Electrophoresis－(DGGE)
Purpose:Denaturing gradient gels are used to detect non-RFLP polymorphisms. The small (200-700 bp) genomic restriction fragments are run on a low to h
Electrophoresis－of－PCR－products－with－Sunrise－gel－apparatus
Electrophoresis of PCR products with Life Technologies Sunrise gel apparatusGel: In a 500 ml Pyrex® glass bottle, add:Agarose:3 gH2O270 mls10X TA30 ml
EGel®－CloneWell－Agarose－Gels
实验概要Instructions are provided below for using the E-Gel®CloneWell pre-cast agarose gels with the E-Gel® iBase™ Power System. For detailed instructio
琼脂扩散试验（gel－diffusion－test）
可溶性抗原与相应抗体在半固体琼脂凝胶内扩散，二者相遇，在比例合适处形成白色沉淀。基本类型有单向扩散和双向扩散两种。 （一）双向琼脂扩散试验（Double immunodiffusion test） 抗原和抗体加到琼脂板上相对应的孔中，两者各自向四周扩散，如两者相对应，浓度比例合适，则经一定时间后，在
2D－Polyacrylamide－Gel－Electrophoresis
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
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
Optimized－Fibrin－Gel－Bead－Assay－for－the－Study－of－Angiogenesis
PREPARING CELLSBring up HUVEC and fibroblasts in M199/10% FBS/Pen-Strep (1:100) 1-2 days before beading.Switch medium to EGM-2 (Clonetics) the day bef
凝胶过滤(gel－filtration,GF)的应用
1.生物大分子的纯化凝胶过滤是依据分子量的不同来进行分离的，由于它的这一分离特性，以及它具有简单、方便、不改变样品生物学活性等优点，使得凝胶过滤成为分离纯化生物大分子的一种重要手段，尤其是对于一些大小不同，但理化性质相似的分子，用其它方法较难分开，而凝胶过滤无疑是一种合适的方法。例如对于不同聚合程度
Isoelectric－Focussing－of－Membrane－Proteins－by－Slab－Gel－Method
REFERENCE: Ames, G.F.L. and Nikaido, H. 1976. Biochemistry. 15:616-623.MATERIALS:Gel solution:1.05 gacrylamide0.032 gbis-acrylamide8.25 gurea6.5 mldis
胶体电泳法（gel－lectrophoresis）方法步骤
SDS 平板胶片铸造︰1） 整理出两组玻璃片及白板铸胶组合，先用酒精拭净，并选择合适的间隔条（0.75 mm） 组合起来。请熟悉铸胶三明治组合的正确组装方式，以免灌入的胶液漏出来。2） 三明治组合后直立站好，准备所要浓度的SDS 胶体溶液如表4.2. 两片0.75mm 厚的平板胶片约需10 mL 分
2－Dimensional－Gel－Electrophoretic－Analysis－for－Chicken－Egg
Overview This protocol is a detail description of the procedure in performing 2D gel electrophoresis for illustrating the protein profile of the w
DNA的凝胶电泳（gel－electrophoresis）
一、原理琼脂糖或聚丙烯酰胺凝胶是分离和纯化DNA片段的标准方法。聚丙烯酰胺凝胶电泳适用于分离小分子的核酸；琼脂糖凝胶孔径较大，被应用于大分子核酸的分离和纯化。在一定浓度的琼脂糖凝胶介质中，DNA分子的电泳迁移率与其分子量的常用对数成反比。当用低浓度的荧光嵌入染料溴化乙啶（EB）染色，在紫外光下至少可
QUALITATIVE－ANALYSIS－OF－DNA－FRAGMENTATION－BY－AGAROSE－GEL－ELECTROPHORESIS
1. IntroductionNuclear morphology changes characteristic of apoptosis appear within the cell together with a distinctive biochemical event: the endonu