SDS-PAGE polyacrylamide gel

One of the most widely used forms of gel electrophoresis is known as sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Polyacrylamide gel has several advantages that account for its extensive use. It has minimal adsorptive properties and produces a negligible electroosmotic effect (Hjelmeland and Chrambach 1981). In identity tests, for the determination of molecular weight, SDS-PAGE has been shown to be an appropriate, fast, and easy method that is often used in quality control laboratories. The use of SDS-PAGE followed by a densitometric analysis, such as MS, is a helpful technique for the determination of peptide or... 

Abbreviations used Fi Fq, H+-translocating ATP synthases from mitochondria, E. coli plasma membranes, plasma membranes of the thermophilic bacterium PS3 and chloroplast membranes CFi, chloroplast coupling factor one ATPase, CFiE. C. 3.6.1.3 SDS-PAGE, polyacrylamide gel electrophoresis in the presence of sodium dodecyl... 

SDS-PAGE - Polyacrylamide Gel Electrophoresis in Sodium Dodecyl Sulfate SNP - Specific Nuclear Protein... 

SDS-PAGE SDS (lauryl-sulfate)-PAGE (polyacrylamide gel electrophoresis)... 

A, Absorption chi, chlorophyll car, carotenoid EET, excitonic energy transfer EF, exoplasmic fracture face EM, electron microscopy FWHM, full width at half maximum lEF, Isoelectric Focusing, LD, linear dichroism LHC, light harvesting complex PAGE, polyacrylamide gel electophoresis PF, protoplasmic fracture face PS, photosystem RC, reaction centre SDS, sodium dodecyl sulphate SSTT, single step transfer time. 

Abbreviations BSA, bovine serum albumin Chi, chlorophyll DCPIP, 2, 6-dichlorophenol indophenol DMBQ, 2, 6-dimethyl-l, 4-benzoquinone DTT, dithiothreitol Fd, ferredoxin Hepes, N-2-hydroxyethyl-piperazine N -2-ethane sulphonate LDAO, lauryldimethyl amine oxide Mes, 4-morpholine ethane sulphonic acid MV, methyl viologen PAGE, polyacrylamide gel electrophoresis PRuK, phosphoribulokinase PS, photosystem Ru5P, ribulose 5-phosphate SDS, sodium dodecyl sulphate TMPD, N, N, N, N -tetramethyl paraphenylene diamine TRX. thioredoxin... 

SDS = sodium dodecyl sulfate [MefCHyl.oCHjOSO O Na ]. also known as sodium lauryl sulfaie. is a common component ol household detergents and shampoos PAGE = polyacrylamide gel electrophoresis... 

Microtubule-associated proteins bind to microtubules in vivo and subserve a number of functions including the promotion of microtubule assembly and bundling, chemomechanical force generation, and the attachment of microtubules to transport vesicles and organelles (Olmsted, 1986). Tubulin purified from brain tissue by repeated polymerization-depolymerization contains up to 20% MAPs. The latter can be dissociated from tubulin by ion-exchange chromatography. The MAPs from brain can be resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). 

The protein was purified by a dialysis procedure, denatured and analysed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Western blotting indicated that the protein of interest consisted of two components, one of which increased in concentration as the purification proceeded. The authors initially suggested that this could be due to the presence of a number of species produced by modification of the amino acid side-chains, for example, by glyco-sylation, or by modification of the C- or N- terminus. 

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) An electrophoretic technique used for the separation of proteins. 

The number of different proteins in a membrane varies from less than a dozen in the sarcoplasmic reticulum to over 100 in the plasma membrane. Most membrane proteins can be separated from one another using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), a technique that has revolutionized their study. In the absence of SDS, few membrane proteins would remain soluble during electrophoresis. Proteins are the major functional molecules of membranes and consist of enzymes, pumps and channels, structural components, antigens (eg, for histocompatibility), and receptors for various molecules. Because every membrane possesses a different complement of proteins, there is no such thing as a typical membrane structure. The enzymatic properties of several different membranes are shown in Table 41-2. 

SDS-PAGE of purified CinnAE (10% polyacrylamide gel). Samples in lanes as follows Lane 1 6 /tg CinnAE after anion-exchange chromatography lane 2, high molecular weight markers from Sigma. 

Figure 3. SDS-PAGE and in situ pectinase activity on pectin and polygalacturonic acid-agarose overlays of culture filtrates of Aspergillus niger N-402 (upper panel) and Aspergillus FP-180 (lower panel) at 2.5, 3.5, 5.5 and 6.5 pHi (Lanes a, b, c, and d, respectively). Electrophoresis on 10% acrylamide slab gel (14 X 8 cm) in the presence of SDS was according to Laemmli (6), run at 30 mA constant current for 2 hours. Crude cell-free samples were concentrated by lyophilization, dialyzed, boiled with sample buffer by 60 sec. and applied to each well. Polyacrylamide gel and overlays were incubated overnight with 0.17 acetate buffer at room temperature.

Prokaryotic cells express hundreds to thousands of proteins while higher eukaryotes express thousands to tens of thousands of proteins at any given time. If these proteins are to be individually identified and characterized, they must be efficiently fractionated. One-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) has typically been use to study protein mixtures of <100 proteins. Onedimensional electrophoresis is useful because nearly all proteins are soluble in SDS, molecules ranging from approximately 10,000 to 300,000 molecular weight can be resolved, and extremely basic or acidic proteins can be visualized. The major disadvantage to one-dimensional gels is that they are not suitable for complex mixtures such as proteins from whole cell lysates. 

First Dimension Optimization After the second-dimension separation has been developed, the first-dimension flow rate is determined. This includes selecting a first-dimension column diameter to work at the flow rate selected. We illustrate the selection process with an application that addresses a column method for proteins that functions as a replacement for planar 2D gel electrophoresis (2DGE) within a narrow molecular weight and p/range. In the planar experiment, isoelectric focusing is performed in the first dimension and sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS/PAGE) in the second dimension. 

Poehling reported a microscale two-dimensional gel electrophoresis system 25 years ago (Poehling and Neuhogg, 1980 Neuhoff, 2000). In that system, separation in the IEF dimension was performed in thin, gel-filled tubes. After IEF, the gel was transferred to a 3 cm x 3.5 cm polyacrylamide gel, where proteins were separated by SDS-PAGE. Several hundred components were resolved from a few micrograms of protein homogenate. 

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