Two-dimensional gel electrophoresis of the membrane-bound protein complexes, including photosystem I, of thylakoid membranes in the presence of sodium oligooxyethylene alkyl ether sulfate/dimethyl dodecylamine oxide and sodium dodecyl sulfate

1991 ◽  
Vol 22 (3) ◽  
pp. 253-260 ◽  
Author(s):  
Toru Hisabori ◽  
Kazuhito Inoue ◽  
Yasuhiro Akabane ◽  
Sunao Iwakami ◽  
Katsushi Manabe
Keyword(s):  
Sodium Dodecyl Sulfate ◽  
Gel Electrophoresis ◽  
Protein Complexes ◽  
Sodium Dodecyl ◽  
Thylakoid Membranes ◽  
Alkyl Ether ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
Ether Sulfate ◽  
Membrane Bound

Fractionation of nucleolar proteins by two-dimensional gel electrophoresis

1976 ◽  
Vol 54 (1) ◽  
pp. 9-14 ◽  
Author(s):  
G. Jackowski ◽  
D. Suria ◽  
C. C. Liew
Keyword(s):  
Sodium Dodecyl Sulfate ◽  
Gel Electrophoresis ◽  
Guanidine Hydrochloride ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
Nucleolar Proteins ◽  
Dodecyl Sulfate ◽  
Ph Range

Isolation of nucleolar proteins was obtained by dissociation in the presence of urea – guanidine hydrochloride, followed by high-speed centrifugation to remove nucleic acids. At least 31 fractions of nucleolar proteins were detected by isoelectrofocusing gel electrophoresis in the pH range 3.5–10. Following two-dimensional gel electrophoresis on sodium dodecyl sulfate – polyacrylamide slab gels, more than 100 components of nucleolar proteins were identified. Two-thirds of nucleolar proteins were located in the pH range 5–8 following isoelectrofocusing. The molecular weights of these classes of proteins were shown to be mostly 30 000 – 70 000 by sodium dodecyl sulfate – polyacrylamide gel electrophoresis.

Comparative study of six methods of protein extraction for two-dimensional gel electrophoresis of proteomic profiling in poplar stems

2013 ◽  
Vol 93 (5) ◽  
pp. 895-901
Author(s):  
Wassim Azri ◽  
Amel Ennajah ◽  
Mai Jing
Keyword(s):  
Sodium Dodecyl Sulfate ◽  
Comparative Study ◽  
Gel Electrophoresis ◽  
Protein Extraction ◽  
Sodium Dodecyl ◽  
Protein Detection ◽  
Two Dimensional ◽  
Proteomic Profiling ◽  
Two Dimensional Gel Electrophoresis ◽  
Dodecyl Sulfate

Azri, W., Ennajah, A. and Jing, M. 2013. Comparative study of six methods of protein extraction for two-dimensional gel electrophoresis of proteomic profiling in poplar stems. Can. J. Plant Sci. 93: 895–901. Protein extraction is a crucial step in two-dimensional gel electrophoresis (2-DE) analysis of proteins, since it can have significant impact on both the quantity and the quality of protein detection. The present study is a comparison between six previously published protocols of protein extraction (A, B, C, D, E, and F) aiming to determine a suitable method to extract total proteins from poplar stems, a recalcitrant plant tissue. The obtained results revealed that method F (optimized method B), combining detergents (CHAPS, Triton X-100, and low sodium dodecyl sulfate amounts) and chaotropes (thiourea and urea), gave the best solution for the problem of protein solubilization. Method F enabled the detection of more than 300 spots reproducible on the 2-DE gel with pH 4–7 immobilized pH gradient strips and 12% sodium dodecyl sulfate–polyacrylamide gel electrophoresis, using silver staining. Our results suggest that Method F is expected to have excellent applications in proteomic studies of poplar stems.

Reproducibility and quality assurance of two-dimensional gel electrophoresis of serum specimens.

1982 ◽  
Vol 28 (4) ◽  
pp. 908-914 ◽  
Author(s):  
R P Tracy ◽  
R M Currie ◽  
D S Young
Keyword(s):  
Quality Assurance ◽  
Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Two Dimensional ◽  
Protein Loss ◽  
Serum Samples ◽  
Silver Stain ◽  
Two Dimensional Gel Electrophoresis ◽  
Report Quality ◽  
Wide Range

Abstract Currently we are using two different ISO-DALT two-dimensional gel electrophoresis systems, designated MC-Iso 1 and MC-Iso 2, for the analysis of serum and plasma samples. Here we report quality-assurance data for both of these systems. CV values for the slopes of the pH gradient (ISO dimension) are 5.6% of less; CV values for the slopes of the molecular-mass curves (log Mr vs relative mobility in the DALT dimension) are 3.4% or less. We examined the various steps of the analysis in detail for reproducibility and protein loss, using radiolabeled albumin, alpha 2-macroglobulin, and beta 2-microglobulin. Generally, in the first dimension, less protein enters the MC-Iso 2 gels (our routine system in which silver stain is used) than enters the MC-Iso 1 gels (our wide-range system for myeloma serum samples, in which the gel is stained with Coomassie Blue), on the average, 87% as much. The CV at this stage for both systems is 5--8%. During equilibration, considerable amounts of protein are lost (approximately 30% in 10 min) from the ISO gel, and the reproducibility is also decreased. Resolution in the DALT dimension has, in most cases, little or no effect on either recovery or reproducibility. Overall, for most proteins expected to appear in an ISO gel of a given pH range, approximately 50--60% of the starting material may be expected to reside in the sodium dodecyl sulfate slab gel, under our conditions. The two most important variables affecting recovery are the concentration of the NaOH (used as catholyte) and the pH of the starting sample. The overall CV for the process is between 8 and 12%.

Variation in goat fibre protein

1989 ◽  
Vol 40 (3) ◽  
pp. 675 ◽  
Author(s):  
DJ Tucker ◽  
AHF Hudson ◽  
A Laudani ◽  
RC Marshall ◽  
DE Rivett
Keyword(s):  
Molecular Weight ◽  
Sodium Dodecyl Sulfate ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Apparent Molecular Weight ◽  
Wool Fibre ◽  
Two Dimensional ◽  
Protein Patterns ◽  
Cashmere Goats

The proteins from a range of cashmere, mohair, angoratcashmere crossbred and wool fibre samples were extracted at pH 8 with 8 M urea containing dithiothreitol, and were then radiolabelled by S-carboxymethylation using iodo(2-14C) acetate. The proteins from each sample were examined by two dimensional polyacrylamide gel electrophoresis in which the separation in the first dimension was according to charge at pH 8.9 and in the second dimension according to apparent molecular weight in the presence of sodium dodecyl sulfate. After electrophoresis the proteins were detected by fluorography. Protein differences in keratin samples from some individual goats existed, although the overall protein patterns were similar. None of the differences were consistent with any one goat fibre type. The protein patterns obtained for fibre samples from individual cashmere goats showed some differences when compared to those found for commercial blends from the same country of origin, indicating that blending can mask any animal-to-animal variation. While the electrophoretic technique does not unequivocally distinguish between cashmere, mohair and angora/cashmere crossbred fibres it does differentiate between wool and goat fibres.

scholarly journals Analysis of H-2 and Ia molecules by two-dimensional gel electrophoresis.

1977 ◽  
Vol 146 (5) ◽  
pp. 1261-1279 ◽  
Author(s):  
P P Jones
Keyword(s):  
Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Surface Proteins ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
Cell Surface Proteins ◽  
Ia Antigens ◽  
Polypeptide Chains ◽  
Kinetics Of ◽  
Electrophoresis Technique

Mouse lymphocyte H-2 and Ia glycoproteins have been analyzed with a two-dimensional (2-D) acrylamide gel electrophoresis technique, in which proteins are separated first according to their charge in isoelectrofocusing gels and then according to their size in sodium dodecyl sulfate gels. Individual polypeptide chains from radiolabeled cells are resolved as discrete spots on autoradiograms of the gels, forming patterns which are characteristic of the proteins in the sample. 2-D gels of H-2K, H-2D, and Ia glycoproteins immunoprecipitated from 35S-methionine-labeled cells reveal that these proteins exist in the cells as complex arrays of molecules heterogeneous in both size and charge. Lactoperoxidase-catalyzed radioiodination of lymphocyte surfaces labels only subsets of the total H-2 and Ia molecules with 125I, indicating that some of the molecules may represent cytoplasmic precursors of the cell surface proteins. This theory is supported by the kinetics of labeling of various spots in 35S-methionine pulse-chase experiments. The 2-D gel patterns obtained for both H-2 and Ia antigens have also been shown to be haplotype-specific and independent of the genetic background.

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