Investigation of complex protein patterns of latex proteins of various Euphorbiaceae Juss. by two dimensional gel electrophoresis

Abstract Using a combination of two-dimensional gel electrophoresis (2DE), silver staining, and computerized densitometry, we studied protein patterns in human cerebral cortex: normal fresh-frozen, fresh-frozen but previously irradiated, and post-mortem. The relative molecular mass of the resolved proteins ranged from 14 400 to 100 000, the isoelectric points from 4.75 to 7.0. The pattern of proteins (six of them identified) was essentially the same for all three groups. However, computerized densitometry demonstrated significant alterations in the density of several spots in the irradiated and postmortem groups as compared with the normal controls. Irradiated cortex showed statistically significant changes in only six spots (three increased and three decreased in density); postmortem material showed 20 altered spots (16 diminished and four increased). Evidently normal human cerebral cortex has a consistent protein pattern on 2DE, which is quantitatively (but not qualitatively) altered in irradiated and postmortem material. These findings provide a point of reference against which proteins from abnormal brain material can be compared, both qualitatively and quantitatively.

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Induction of Heat-Shock Protein (hsp) by Moxibustion

The American Journal of Chinese Medicine ◽

10.1142/s0192415x95000390 ◽

1995 ◽

Vol 23 (03n04) ◽

pp. 327-330 ◽

Cited By ~ 13

Author(s):

Kazuko Kobayashi

Keyword(s):

Heat Shock ◽

Heat Shock Protein ◽

Gel Electrophoresis ◽

Two Dimensional ◽

Hsp 70 ◽

Protein Patterns ◽

Two Dimensional Gel Electrophoresis ◽

Hip Muscle ◽

And Control ◽

Muscular Tissues

Rats were treated by moxibustion at the point of hip muscle, and intramuscular temperature was kept at 40°C for 15 minutes. The rats were sacrificed under deep anesthesia and the muscular tissues were excised immediately, three hours and 24 hours after stimulation. Proteins were extracted from the homogenized and centrifuged tissues of the stimulated rats and control rats. Two-dimensional gel electrophoresis of the proteins was carried out. Heat-shock protein (hsp) with molecular weight of 70,000 (hsp 70), 85,000 (hsp 85) and 100,000 (hsp 100) was detected in rats sacrificed three hours after the stimulation by moxibustion. Protein patterns were analyzed and the ratios of the hsps were obtained.

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Protein patterns of human nasal and bronchoalveolar lavage fluids analyzed with two-dimensional gel electrophoresis

Electrophoresis ◽

10.1002/elps.1150191828 ◽

1998 ◽

Vol 19 (18) ◽

pp. 3222-3229 ◽

Cited By ~ 53

Author(s):

Mats Lindahl ◽

Bengt Ståhlbom ◽

Jesper Svartz ◽

Christer Tagesson

Keyword(s):

Bronchoalveolar Lavage ◽

Gel Electrophoresis ◽

Two Dimensional ◽

Protein Patterns ◽

Two Dimensional Gel Electrophoresis

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Combination of strong anion exchange liquid chromatography with microchip capillary electrophoresis sodium dodecyl sulfate for rapid two-dimensional separations of complex protein mixtures

Analytical and Bioanalytical Chemistry ◽

10.1007/s00216-021-03797-4 ◽

2021 ◽

Author(s):

Holger Zagst ◽

Christin Elgert ◽

Sönke Behrends ◽

Hermann Wätzig

Keyword(s):

Capillary Electrophoresis ◽

Liquid Chromatography ◽

Gel Electrophoresis ◽

Sodium Dodecyl ◽

Soluble Guanylyl Cyclase ◽

Two Dimensional ◽

Microchip Capillary Electrophoresis ◽

Two Dimensional Gel Electrophoresis ◽

Complex Protein ◽

Strong Anion Exchange

AbstractTwo-dimensional separations provide a simple way to increase the resolution and peak capacity of complex protein separations. The feasibility of a recently developed instrumental approach for two-dimensional separations of proteins was evaluated. The approach is based on the general principle of two-dimensional gel electrophoresis. In the first dimension, semi-preparative strong anion exchange high-performance liquid chromatography is utilized and fractions are collected by means of a fraction collector. They are subsequently analyzed in the second dimension with microchip capillary electrophoresis sodium dodecyl sulfate. Microchip capillary electrophoresis provides the necessary speed (approximately 1 min/fraction) for short analysis. In this study, three different samples were investigated. Different constructs of soluble guanylyl cyclase were expressed in Sf9-cells using the baculovirus expression system. Cell lysates were analyzed and the resulting separations were compared. In our experimental setup, the soluble guanylyl cyclase was identified among hundreds of other proteins in these cell lysates, indicating its potential for screening, process control, or analysis. The results were validated by immunoblotting. Samples from Chinese hamster ovary cell culture before and after a purification step were investigated and approximately 9% less impurities could be observed. The separation patterns obtained for human plasma are closely similar to patterns obtained with two-dimensional gel electrophoresis and a total of 218 peaks could be observed. Overall, the approach was well applicable to all samples and, based on these results, further directions for improvements were identified. Graphical abstract .

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Two-dimensional protein patterns of tomato (Lycopersicon esculentum Mill.) seeds; effects of isolation procedure and imbibition

Seed Science Research ◽

10.1017/s0960258500002300 ◽

1994 ◽

Vol 4 (3) ◽

pp. 275-283 ◽

Cited By ~ 5

Author(s):

J. H. W. Bergervoet ◽

H. L. Kraak ◽

C. H. R. De Vos ◽

R. J. Bino

Keyword(s):

Lycopersicon Esculentum ◽

Gel Electrophoresis ◽

Protein Pattern ◽

Two Dimensional ◽

Protein Patterns ◽

Two Dimensional Gel Electrophoresis ◽

General Picture ◽

Isolation Methods ◽

Lysis Buffer

AbstractA computer-aided comparison of tomato (Lycopersicon esculentum Mill.) seed protein patterns, obtained after two-dimensional gel electrophoresis, was made for three different extraction procedures: TCA acetone/lysis buffer, lysis buffer only and modified Laemmli/lysis buffer. Comparison of the isolation methods showed that about half of the amount of proteins detected was common in each method. Also, proteins specific to some isolation methods were detected. Protein synthesis during imbibition was monitored using 35S-methionine. After labelling the proteins were extracted using TCA acetone/lysis buffer. Following two-dimensional gel electrophoresis the gels were first silver stained, to give a general picture of all proteins present in the seed, then the gels were exposed to a film for autoradiography. Comparison of the in vivo-synthesized protein patterns and the silver-stained proteins revealed that from day 0 to day 1 the protein pattern was changed but the total number of different spots was similar. After 1 day of imbibition, the number of protein spots increased greatly and the protein pattern changed again.

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