Electrophoresis pattern

Fig. 3.161. (A) Zone electrophoresis patterns of FITC-labelled transferrin samples by fluorescence detection. The unbound dye (providing a main peak and several minor ones) was not removed from the samples. Experimental conditions background electrolyte, 100 mM borate buffer, pH 8.3 voltage, 20 kV capillary 59 cm (effective length 41 cm) X 75 pm i.d. injection of samples 100 mbar x s 20°C detection with fluorescence detector (240 - 400 nm, broadband excitation filter and a 495 nm cut-off emmision filter). The reaction was left to continue for 20 h, and the reaction mixtures contained 13 pm (1 mg/ml) Tf and (a) 0.01 mM FITC, (b) 0.1 mM FITC, and 1 mM FITC. (B) Zone electrophoresis patterns of an FITC-labelled transferrin sample by simultaneous fluorescence (upper trace, left axis) and UV detection (lower trace, right axis). The unbound dye shows several peaks with both detections. Experimental conditions background electrolyte, 100 mM borate buffer, pH 8.3 voltage, 20 kV capillary 59 cm (effective length fluorescence 41 cm, UV 50.5 cm) X 75 pm i.d. injection of samples 100 mbar X s 20°C detection with fluorescence detector (240 - 400 nm, broadband excitation filter and a 495 nm cut off emmision filter). The reaction was left to continue for 20 h, and the reaction mixtures contained 6.5 pm (0.5 mg/ml) Tf and 0.1 mM FITC. Reprinted with permission from T. Konecsni et al. [199].

Fig. 5.5 Electrophoresis pattern of proteins in the allantoic fluid of chicken embryos. Lanes 1-3 intact fluid lanes 4-6 fluid after 6h of irradiation...

Fig. 5.6 Electrophoresis pattern of proteins of blood serum. Lanes 1-3 intact serum, lanes 4-6 serum after 6 h of irradiation (See Color Plates)...

To elucidate some enzymatic characteristics of the isolated laccases I, II, and III, substrate specificities for several simple phenols, electrophoresis patterns, ultraviolet spectra, electron spin resonance spectra, copper content, and immunological similarities were investigated. Tyrosine, tannic acid, g c acid, hydroquinone, catechol, pyrogallol, p-cresol, homocatechol, a-naphthol, -naphthol, p-phenylenediamine, and p-benzoquinone as substrates. No differences in the specificities of these substrates was found. The UV spectra for the laccases under stucfy are shown in Figure 4. Laccase III displays three adsorption bands (280, 405, and 600nm), laccase II shows one band 280nm), and laccase I shows two bands (280 and 405 nm). These data appear to indicate differences in chemical structure. The results of the copper content analysis (10) and two-dimensional electrophoresis also indicate that these fractions are completely different proteins (10), Therefore, we may expect differences in substrate specificities between the three laccase fractions for more lignin-like substrates, yet no difference for some simple phenolic substrates. 

Fig. 4.1.9 Electrophoresis pattern of different types of MPS (indicated beneath the corresponding band). Standards (STD) and normal controls (N) were also run on each gel. GAGs are labeled on the left side of the figure. The picture was kindly provided by Dr. E Buerger, Metabolic Center Heidelberg, Germany. GM1 GM1-gangliosidosis, MSD multiple sulfatase deficiency ML II mucolipidosis II, LZ loading zone of the gel...

The results of the lipoprotein electrophoresis have to be interpreted in the context of other lipid parameters, like plasma total cholesterol and triglyceride levels. Patients with normal cholesterol and triglyceride values may sometimes show electrophoresis patterns that resemble pathologic patterns but should not be classified as such. For untreated type III patients, plasma total cholesterol levels should range from 7.5 to 13.0 mmol/1 and triglycerides from 3.5 to 10.5 mmol/1. The presence of a broad-ji-band in the absence of hyperlipidemia excludes familial dysbetalipoproteinemia (type III). 

FIG. 12.12 Electrophoresis patterns for human serum (a) schematic of schlieren profiles and (b) semilog plot of protein molecular weight versus electrophoretic mobility for particles electro-phoresed on cross-linked polyacrylamide. (Reprinted with permission from K. Weber and M. Osborn, J. Biol. Chem., 244, 4404 (1969).)... 

GL Lookhart, Y Pomeranz. Gliadin high-performance liquid chromatography and polyacrylamide gel electrophoresis patterns of wheat grown with fertilizer treatments in the United States and Australia on sulfur-deficient soils. Cereal Chem 62 227-229, 1985. 

Southern blotting. A method for detecting a specific DNA restriction fragment, developed by Edward Southern. DNA from a gel electrophoresis pattern is blotted onto nitrocellulose paper then the DNA is denatured and fixed on the paper. Subsequently, the pattern of specific sequences in the Southern blot can be determined by hybridization to a suitable probe and autoradiography. A northern blot is similar, except that RNA is blotted instead onto the nitrocellulose paper. 

FIGURE 16.20 (a) A normal electrophoresis pattern of blood serum, (b) An abnormal pattern, with elevated y-globulin, indicating the possibility of liver disease, collagen disorder, or infection. 

Scheler, C., Muller, E. C., Stahl, J., Muller-Werdan, U, Salnikow J, and Jungblut, P. (1997) Identification and characterization of heat shock protein 27 protein species in human myocardial two-dimensional electrophoresis patterns. Identification and characterization of heat shock protein 27 protein species in human myocardial two-dimensional electrophoresis patterns. Electrophoresis 18, 2823-2831. 

When the area of the membrane containing the protein of interest is known, another procedure, described by Lindahl (91), can be used. Only a limited area of the nitrocellulose containing the proteins is cut out and incubated with antibodies. The rest of the membrane is stained with one of the methods described in Note 10 (a method not compatible with immunodetection, but much more sensitive than Ponceau S, can be used). Thus the protein spots can be located in the 2D electrophoresis pattern, matched with a corresponding silver stained gel and translated into the protein pattern. This method also allows a considerable saving of antibodies. 

Zeindl-Eberhrt, E., Jungblut, P. R., and Rabes, H. M. (1997) A new method to assign immunodetected spots in the complex two-dimensional electrophoresis pattern. Electrophoresis 18, 799-801. 

Simon, N., Radioactive gold in filter paper electrophoresis patterns of plasma. Science 119, 95 (1954). 

Figure 7.4 (a) Polyacrylamide gel electrophoresis patterns forthe hydrolysis... 

Figure 7.10 Agarose gel electrophoresis patterns for Ce(iv)/EDTA-induced site-selective hydrolysis of double-stranded DNA by Ce(iv)/EDTA and pcPNA additives. Bands were detected by staining with GelStar. (a) Site-selective hydrolysis of linearized PBR322 using pcPNAs. Lane 1, control lane 2, Ce(iv)/...

Fig- ( ) Gel electrophoresis patterns of immune serum and affinity purified antibodies [A], Density gradient ultracentrifugation (B), top pattern glucose oxidase, lower pattern mesquite antibody. 

Figure 4.15 Moving boundary electrophoresis pattern of normal human serum (Reproduced with permission from Bezkorovainy A. Basic Protein Chemistry. Springfield, IL Thomas, p. 20, 1970.)...

Figure 6-4. Sodium dodecyl sulfate polyacrylamide gel electrophoresis pattern of normal red cell membrane proteins with Coomassie blue staining.

Outline the process of DNA sequencing, and deduce a DNA sequence from an electrophoresis pattern. 

Dunn, M. )., Jungblut, P. R. (1999). Gomparison of two-dimensional electrophoresis patterns of heat shock protein Hsp27 species in normal and cardio-myopathic hearts. Electrophoresis 20, 3623-3628. 

Problem 28.18 Sketch what ynu would expect the gel electrophoresis pattern to kK like if the DNA segment In Frobtem 2. 17 were itequenccd. 

Fig. 6. Moving boundary electrophoresis patterns at pH 4.5 of high-sulfur fractions SCMKB from wools of various breeds of sheep (Gillespie, unpublished observations, 1964).

The protein components of a membrane can be readily visualized by SDS-polyacrylamide gel electrophoresis. As discussed earlier (Section 4.1.4). the electrophoretic mobility of many proteins in SDS-containing gels depends on the mass rather than on the net charge of the protein. The gel-electrophoresis patterns of three membranes—the plasma membrane of erythrocytes, the photoreceptor membrane of retinal rod cells, and the sarcoplasmic reticulum membrane of muscle—are shown in Figure 12.16. It is evident that each of these three membranes contains many proteins but has a distinct protein composition. In general, membranes performing different functions contain different repertoires of proteins. 

Once separated, the locations of the DNA cleavage products are detected by exposing the gel to a photographic plate, a process called autoradiography. Each radioactive end piece containing a label appears as a dark band on the photographic plate, but nonradioactive pieces from the middle of the chain aren t seen. The gel electrophoresis pattern shown in Figure 28.14 would be obtained in our hypothetical example. 

Representation of a gel electrophoresis pattern. The products of the four cleavage experiments are placed at the top of the gel, and a voltage is applied between top and bottom. Smaller products migrate along the gel at a faster rate and thus appear at the bottom. 

Problem 28.19 Finish assigning the sequence to the gel electrophoresis pattern shown in Figure 28.14. 

AAT is the major constituent of the tti-glohulin band on routine clinical serum electrophoresis. The two other relatively high concentration tt]-globulins, AAG and a-lipoprotein, do not stain well with peptide stains because of their high contents of CHO and lipid, respectively. Some genetic variants of AAT may be detectable by visual examination of the electrophoresis pattern because of altered mobility or decreased concentration. 

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