Separation of ferritin dimer, trimer and tetramer by preparative polyacrylamide gel electrophoresis and their identification by electron microscopy

1991 ◽  
Vol 19 (2) ◽  
pp. 218S-218S
Author(s):  
SHEIKH A. SAEED ◽  
TOM R.C. BOYDE
Keyword(s):  
Electron Microscopy ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel

Intracellular fibres in cultured cells: analysis by scanning and transmission electron microscopy and by SDS-polyacrylamide gel electrophoresis

1978 ◽  
Vol 31 (1) ◽  
pp. 369-392
Author(s):  
J.A. Trotter ◽  
B.A. Foerder ◽  
J.M. Keller
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Dimensional Structure ◽  
3T3 Cells ◽  
Transmission Electron ◽  
Ionic Detergent ◽  
Over The Top

The 3-dimensional structure of the fibrous cytoskeleton of 3T3 cells was examined by scanning electron microscopy of cells extracted with the non-ionic detergent Triton X-100. Detergent-extracted cells consist of the nucleus and an extensive system of fibres, the largest of which correspond to stress fibres visible by phase-contrast microscopy. The system of fibres, which is coterminous with the borders of the native cell, remains firmly adherent to the substratum. The major fibres branch into smaller fibrils which appear to end by ravelling out into fine filaments that constitute a matted network in a plane very close to that of the substratum. In the nuclear region all the major fibres pass over the top of the nucleus, where they may also branch into a system of fine fibrils. Thin-section transmission electron microscopy in conjunction with heavy meromyosin treatment of extracted cells shows the fibres to be composed of native F-actin. Intermediate filaments are also present, and are prominent in the matted network, together with actin filaments. The major proteins of the residue are identified by SDS-polyacrylamide gel electrophoresis as actin, a 56000 Dalton peptide, and histones. Also present are myosin heavy chain, peptides of 225,000 and 250,000, and minor bands at 60,000 and 94,000 Daltons. The non-ionic detergent extracts 70% of the cellular protein, including 50% of the actin and 75% of the myosin. The Triton-insoluble fraction of 3T3 cells appears to constitute, in addition to the nucleus, a stable cytoskeletal system, composed largely of contractile proteins and 10-nm filaments, which functions in maintenance of cell shape, in substratum adhesion, and in positioning the nucleus within the cell.

scholarly journals Comparasion of polyacrylamide gel electrophoresis (PAGE), immuno-electron microscopy (IEM) and enzyme immunoassay (EIA) for the rapid diagnosis of rotavirus infection in children

1983 ◽  
Vol 78 (4) ◽  
pp. 483-490 ◽  
Author(s):  
H. G. Pereira ◽  
R. S. Azeredto ◽  
F. Sutmoller ◽  
J. P. Leite ◽  
V. de Farias ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Enzyme Immunoassay ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Complete Agreement ◽  
Diagnostic Assay ◽  
Negative Results ◽  
Highly Sensitive ◽  
Immuno Electron Microscopy

Detection of rotavirus RNA by polyacrylamide gel electrophoresis (PAGE) proved to be a highly sensitive and rapid diagnostic test. A comparison of this assay with immuno-electron microscopy (IEM) and enzyme immunoassay (EIA) in 245 faeces from children with gastroenteritis revealed complete agreement between the three assays in 238 (97.14%) samples. Among 75 samples positive in at least one of the three assays, negative results were observed in 5 (6.48%) by PAGE, in 6 (6.76%) by EIA and in none by IEM. Silver staining greatly increased the sensitivity of the PAGE assay. We conclude that although IEM remains the most sensitive and rapid rotavirus diagnostic assay, the PAGE technique has many advantages in its favour, including the non-requirement of expensive equipment, the use of only chemically defined reagents and the capacity to distinguish virus subgroup and variants and to detect non-crossreactive rotaviruses which are missed in serological assays.

scholarly journals Antigenic heterogeneity of gonococcal pili.

1975 ◽  
Vol 141 (6) ◽  
pp. 1470-1475 ◽  
Author(s):  
T M Buchanan
Keyword(s):  
Electron Microscopy ◽  
Sodium Dodecyl Sulfate ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Significant Inhibition ◽  
Polyacrylamide Gel ◽  
Dodecyl Sulfate ◽  
Different Strains ◽  
Antigenic Heterogeneity

Pili were isolated from two different strains of gonococci (33 and 2686) and demonstrated to be pure by the criteria of electron microscopy and polyacrylamide gel electrophoresis in sodium dodecyl sulfate. Each purified pili preparation was studied for its ability to inhibit; (a) binding of 124-I-labeled purified 2686 pili by antibody to 2686 pili, and (b) binding of 125-I-labeled purified 33 pili by antibody to 33 pili. In each instance progressive inhibition of binding was produced by homologous pili type, but no significant inhibition was observed using comparable weights of the heterologous pili type. These results indicate that the pili of strains 2686 and 33 are antigenically different.

Ultrastructural localization of specific nucleoprotein fractions

1978 ◽  
Vol 36 (2) ◽  
pp. 204-205
Author(s):  
G. L. Brown
Keyword(s):  
Gel Electrophoresis ◽  
Mouse Liver ◽  
Polyacrylamide Gel Electrophoresis ◽  
Ultrastructural Localization ◽  
Polyacrylamide Gel ◽  
Acid Extraction ◽  
Acid Solutions ◽  
Low Salt ◽  
Liver Nuclei ◽  
Phosphate Groups

Bismuth (Bi) stains nucleoproteins (NPs) by interacting with available amino and primary phosphate groups. These two staining mechanisms are distinguishable by glutaraldehyde crosslinking (Fig. 1,2).Isolated mouse liver nuclei, extracted with salt and acid solutions, fixed in either formaldehyde (form.) or gl utaraldehyde (glut.) and stained with Bi, were viewed to determine the effect of the extractions on Bi stainina. Solubilized NPs were analyzed by SDS-polyacrylamide gel electrophoresis.Extraction with 0.14 M salt does not change the Bi staining characteristics (Fig. 3). 0.34 M salt reduces nucleolar (Nu) staining but has no effect on interchromatinic (IC) staining (Fig. 4). Proteins responsible for Nu and glut.- insensitive IC staining are removed when nuclei are extracted with 0.6 M salt (Fig. 5, 6). Low salt and acid extraction prevents Bi-Nu staining but has no effect on IC staining (Fig. 7). When nuclei are extracted with 0.6 M salt followed by low salt and acid, all Bi-staining components are removed (Fig. 8).

Sign in / Sign up

Export Citation Format

Share Document