EdmanSequencingofProteinsfrom2DGels

The Western blotting/sequencing technique using polyvinylidene difluoride (PVDF) membrane is one of the most popular technique for Edman sequencing. A protein sample is transferred from a 2D polyacrylamide gel electrophoresis (2D-PAGE) gel onto a PVDF membrane by electroblotting. The membrane carrying the protein is directly subjected to protein sequencing. If sequencing fails after a few cycles, the PVDF......阅读全文

Edman-Sequencing-of-Proteins-from-2D-Gels

The Western blotting/sequencing technique using polyvinylidene difluoride (PVDF) membrane is one of the most popular technique for Edman sequencin

NO-WEDGE-Sequencing-Gels

I have been using an alternative to wedge gels that saves acrylamide, cuts gel drying time to 20 min and gives as good or better band squashing at the

DNA-Sequencing-Gels

DNA Sequencing GelsBuffers and gel solutionsLong Ranger: we started using this in early 1995. Great stuff; the best thing is that the gels are not sti

PREPARATION-OF-SEQUENCING-GELS

MATERIALS:2-glass plates1 sharks -tooth comb and spacersWhatman 3 mm paper30 or 40% acrylamide-bis (19:1)10X TBEurea10% ammonium persulfateTEMED60 cc.

peptide-fingerprint-mapping

Polyacrylamide gel electrophoresis is a widely used technique to separate proteins from biological samples. Moreover, the development of two-dimension

Ingel-digestion-of-proteins-for-peptide-fingerprint-mapping

Polyacrylamide gel electrophoresis is a widely used technique to separate proteins from biological samples. Moreover, the development of two-dimension

DNA-Purification-from-Agarose-Gels

1. Separate DNA fragments in an agarose gel cast with 0.5 mg/mL Ethidium bromide. Locate bands with a hand-held long-wave UV lamp.2. Slice the gel wit

2D-凝胶的蛋白-Edman-测序实验

试剂、试剂盒SDS 抽样缓冲液分离胶缓冲液浓缩胶缓冲液SDS-PAGE 电泳缓冲液实验步骤3.1 Cleveland 肽图谱[5]( 1 ) 样品进行 2D-PAGE 分离,然后用考马斯亮蓝(CBB ) 染色,去除含有蛋白质点的凝胶碎片。( 2 ) 用电泳浓缩仪洗脱凝胶中的蛋白质,2W 恒功率洗脱

2D-凝胶的蛋白-Edman-测序实验

试剂、试剂盒:SDS 抽样缓冲液                                                                  分离胶缓冲液                                                              

2D-凝胶的蛋白-Edman-测序实验

试剂、试剂盒 SDS 抽样缓冲液分离胶缓冲液浓缩胶缓冲液SDS-PAGE 电泳缓冲液实验步骤 3.1 Cleveland 肽图谱[5]( 1 ) 样品进行 2D-PAGE 分离,然后用考马斯亮蓝(CBB ) 染色,去除含有蛋白质点的凝胶碎片。( 2 ) 用电泳浓缩仪洗脱凝胶中的蛋白质,2W 恒功率洗

Analysis-of-Proteins-using-Small-Format-2D-Gel-Electrophoresis

Preparation of protein samplesIntracellular virus proteinsThe following method has been developed principally for the analysis of intracellular protei

The-peptide-de-novo-sequencing-from-MS/MS-spectrum

Tandem mass spectrometry (MS/MS) now plays a very important role in protein identification due to its fastness and its high sensitivity.The deriva

Isolation-of-Cell-Wall-Proteins-from-Medicago-sativa-Stems

Plant cell walls are highly dynamic and chemically active components of plant cells. Cell walls consist primarily of polysaccharides, with protein

蛋白质电泳

蛋白质电泳(主要内容如下)One-Dimensional SDS-PAGETwo-Demensional SDS-PAGEProtein Electrophoresis in Agarose Gel Gel StainingRecipesOne-Dimensional SDS-PAGE·      

Preparation-of-Bacterial-Proteins-for-Analysis-by-2DPAGE

The following protocol has been developed for preparing soluble bacterial proteins in a form suitable for analysis by 2D-PAGE. The procedure was princ

Purifying-Large-E.-coli-Restriction-Fragments-from-PulsedField-Gels

DNA PreparationE. coli chromosomal DNA is prepared following the method of Heath et al. ( J. Bacteriol., 174, 1992). Cells are embedded in agarose, th

蛋白组学实验中的常见问题及解答

1,关于使用三氯醋酸沉淀法后蛋白质难于重悬的问题Q:1,Hi !I have been trying to clean and concentrate proteins form eucariotic cell culture medium in order to analyse them in 2

SEMIDRY-ELECTROPHORETlC-TRANSFER-(WESTERN-BLOTS)

 Introduction    After proteins have been separated by electrophoresis, individual protein bands can often be identified by using an antibody that is

DNA抽提

DNA抽提(主要内容如下)·   Working with DNA·   DNA Extraction from Bacteria and Other Organisms·   DNA Extraction from Cell and Tissue·   Mitochondria DNA Isola

RNA-Electrophoresis

Electrophoresis through agarose or polyacrylamide gels is the standard way to separate, identify and purify nucleic acid fragments. The location of th

DNA电泳

DNA电泳(主要内容如下)  Preparation of Agarose Gel and Electrophoresis  Extraction of DNA From Agarose Gel  Extraction of DNA from Acrylamide Gels  DNA Marker 

Lipoprotein-Analysis-Week-2:-Electrophoresis

Lipoprotein Analysis  Week 2: Electrophoresis IntroductionSDS polyacrylamide gel electrophoresis (SDS PAGE) will be used to assess the purification pr

NuPAGE-Gels

NuPAGE GelsA gel electrophoresis system used for SDS-PAGE protein analysis. The gels are made up of Bis-Tris-HCl (pH 6.4) polyacrylamide and are inten

Amino-acid-composition

There has been a recent revival of interest in the use of AA composition for the identification of proteins from 2-D gels. This technique uses the idi

酵母准备

Yeast DNA PreparationYeast Genomic Preparation  (Gottschling Lab)Rapid method for yeast genomic DNA isolation  Yeast DNA Preparation (rapid glass bead

DNA测序

DNA测序(主要内容如下)·         Sequencing Gel Preparation·         Preparation of Templates ·         DNA Sequencing by the Dideoxy Method·         DNA Sequen

非变性胶蛋白电泳

 Section 2.1Nondenaturing Polyacrylamide Gel Electrophoresis of ProteinsJohn M. Walker1. IntroductionSDS-PAGE (Section 2.2) is probably the most commo

Preparation-of-Polyacrylamide-Gels

1. Prepare 20X TBE as:216 g Tris Base110 g Boric Acid80 mL 500 mM EDTA, pH 8.0700 mL ddH2OMix. Bring volume to 1 L. Autoclave.2. Prepare Acrylamide so

DNA-mobility-in-gels

1. Migration of marker dyes in native polyacrylamide non-denaturing gels Gel % Bromophenol blue (BP) Xylene cyanole (XC)   3.5  100 460   5.0

The-E.Z.N.A.®-MagBind™-Dye-terminator-Removal-Procedure

实验概要Excess  unincorporated, nonradioactive label can cause high background  fluorescence in automated sequencing gels. For optimal sequencing  results