Electrophoresis analysis

In protein electrophoresis, a sample is applied to a polyacrylamide gel and its protein components are separated by application of an electric field across the gel. Separation is dependent on the charge and size of the proteins in the sample. Different approaches to this method have been developed to suit a variety of purposes. 

CAUTION Voltages and currents used for electrophoresis are potentially lethal. Use properly shielded, safety-certified equipment. 

10% (w/v) ammonium persulfate (make fresh) Tetramethylethylenediamine(TEMED) 

Protein molecular weight standards appropriate for gel percentage (optional) 

Power supply capable of delivering constant current at voltages 250 V 

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G. Muyzer, E. C. de Waal, and A. G. Uitterlinden, Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for I6S rRNA. Appl. Environ. Microbiol. 59 695 (1993). 

Hurst, W.J. and Martin, R.A. Jr, The quantitative determination of caffeine in beverages using capillary electrophoresis analysis, 21,389-91,1993. 

FIGURE 15.4 Computer record of a two-dimensional capillary electrophoresis analysis of a protein homogenate prepared from ahiopsy obtained from the fundus of a Barrett s esophagus patient. The data were generated hy performing 1 s transfers between capillaries and a 9 s second-dimension separation. The first-dimension separation employed the same buffer as the CSE separation in Fig. 15.1 and the second-dimension separation employed the same buffer as the MECC separation in Fig. 15.1. 

FIGURE 15.10 Landscape image of a two-dimensional capillary electrophoresis analysis of the protein content of a single fixed MCF-7 breast-cancer cell. The data consist of a few high-amplitude components. 

Michels, D.A., Hu, S., Damhrowitz, K.A., Eggertson, M.J., Lauterhach, K., Dovichi, NJ. (2004). Capillary sieving electrophoresis-micellar electrokinetic chromatography fully automated two-dimensional capillary electrophoresis analysis of Deinococcus radiodurans protein homogenate. Electrophoresis 25, 3098-3105. 

Carboni L, Piubelli C, Righetti PG, Jansson B, Domenici E. 2002. Proteomic analysis of rat brain tissue comparison of protocols for two-dimensional gel electrophoresis analysis based on different solubilizing agents. Electrophoresis 23 ... 

Kaale, E., Leonard, S., Van Schepdael, A., Roets, E., and Hoogmartens, J. (2000). Capillary electrophoresis analysis of gentamicin sulphate with UV detection after pre-capillary derivatization with 1,2-phthalic dicarboxaldehyde and mercaptoacetic acid. /. Chromatogr. A 895, 67—79. 

Lurie, I. S., Hays, P. A., and Parker, K. (2004). Capillary electrophoresis analysis of a wide variety of seized drugs using the same capillary with dynamic coatings. Electrophoresis 25, 1580—1591. 

Also known as copper blue. maunt on blu ) moving-boundary electrophoresis analychem A U-tube variation of electrophoresis analysis that uses buffered solution so that all ions of a given species move at the same rate to maintain a sharp, moving front (boundary). miivii) baun-dre i lek-tro-fo re-sos ... 

RC Tim, RA Kautz, BL Karger. Ultratrace analysis of drugs in biological fluids using affinity probe capillary electrophoresis analysis of dorzolamide with fluorescently labeled carbonic anhydrase. Electrophoresis 21 220—226, 2000. 

K Shimura, BL Karger. Affinity probe capillary electrophoresis Analysis of recombinant human growth hormone with a fluorescent-labeled antibody fragment. Anal Chem 66 9-15, 1994. 

An electrophoresis analysis in free solution (CZE), calls for the use of a capillary of 32 cm and with effective length of separation 24.5 cm. The applied voltage is 30 kV. Under the conditions of the experiment the peak of a neutral marker appeared upon the electropherogram at 3 min. 

R. Gillette, and J. V. Sweedler, Capillary Electrophoresis Analysis of NO Synthase Related Metabolites in Single Neurons, Anal. Chem. 1998, 70, 2243. 

Schmitt-Kopplin, P., Garmash, A. V., Kudryavtsev, A. V., Menzinger, F., Perminova, I. V., Hertkorn, N., Freitag, D., Petrosyan, V. S., and Kettrup, A. (2001a). Quantitative and qualitative precision improvements by effective mobility-.scale data transformation in capillary electrophoresis analysis. Electrophoresis 22,77-87. 

Fig. 4. Massive production of green fluorescent protein (GFP) by the continuous-flow cell-free method. Sodium dodecyl sulfide-polyacrylamide gel electrophoresis analysis of GFP produced during 14 d of reaction. mRNA produced by transcription of circular plasmid of Ehime University (pEU) was used for the translation reaction in the dialysis membrane system and was added every 48 h. A 0.1 -pL aliquot of the mixture was run on the gel and protein bands were stained with Coomassie Brilliant Blue. The arrow shows GFP and st designates an authentic GFP band (0.5 pg).

Comparative gel electrophoresis analysis of the 4H junction of U1 snRNA showed the junction adopted a coaxially stacked structure with almost perpendicular axes (Fig. 7.3). This result was very recently confirmed crystallographically (Pomeranz-Krummel et al., 2009). Perhaps the most extensively studied 4H junction in RNA is that of the hairpin ribozyme,... 

Conditions for the Capillary Zone Electrophoresis Analysis of Aspartame... 

Methods to determine the potential biological activity of products obtained through recombinant DNA techniques are of fundamental importance. Despite the existence of numerous physicochemical techniques to characterize the protein product structure and the presence of contaminants, they provide little, if any, information about its biological potency. A bioassay is defined as a functional test, and no physicochemical test can measure the function. However, for some peptide hormones, which are less complex in structure than most cytokines, well defined physicochemical tests may be used to estimate biological activity for instance, the capillary electrophoresis analysis of a protein s isoform content if the specific activity of each one is known. 

Mercier, J.-P., Morin, Ph., Dreux, M. and Tam-bute, A. Capillary electrophoresis analysis of chemical warfare agent breakdown products, J. Chromatogr., A, 741, 279 (1996). 

Lagally ETS, P. C., Mathies, R. A. Monolithic integrated microfluidic DNA amplification n and capillary electrophoresis analysis system. Sens Actuators B 2000 B63 138-146. 

Giavalisco, P., Nordhoff, E., Lehragh, H., Gobom, j., Klose, j., Mueller, M., Egelhoeer, V., Theiss, D., Seitz, H. (2003). Extraction of proteins from plant tissues for two-dimensional electrophoresis analysis A calibration method that simplifies and improves accurate determination of peptide molecular masses by MALDl-TOE MS. Electrophoresis 24, 207-216. 

Figure 15.1 General scheme showing the steps to be followed for the preparation of cytosolic, mitochondrial, and membrane brain proteins and for the enrichment of the cytosolic proteins prior to the 2-D gel electrophoresis analysis.

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