Immunofluorescent Localization of MAPKs in Steedman’s Wax Sections

2014 ◽  
pp. 117-130 ◽  
Author(s):  
Miroslav Ovečka ◽  
Olga Šamajová ◽  
František Baluška ◽  
Jozef Šamaj
Keyword(s):  
Immunofluorescent Localization

Immunocytochemical localization of desmin and vimentin in chick embryonic myocardial cells

1990 ◽  
Vol 48 (3) ◽  
pp. 448-449
Author(s):  
Yukiko Sugi
Keyword(s):  
Sodium Methoxide ◽  
Intermediate Filament Protein ◽  
Chick Embryos ◽  
Immunocytochemical Localization ◽  
Myocardial Cells ◽  
Immunofluorescence Study ◽  
Immunofluorescent Localization ◽  
Rabbit Igg ◽  
Semithin Sections ◽  
Filament Protein

In cultured skeletal muscle cells of chick, one intermediate filament protein, vimentin, is primarily formed and then synthesis of desmin follows. Coexistence of vimentin and desmin has been immunocytochemically confirmed in chick embryonic skeletal musclecells. Immunofluorescent localization of vimentin and desmin has been described in developing myocardial cells of hamster. However, initial localization of desmin and vimentin in early embryonic heart has not been reported in detail. By quick-freeze deep-etch method a loose network of intermediate filaments was revealed to exist surrounding myofibrils. In this report, immunocytochemical localization of desmin and vimentin is visualized in early stages of chick embryonic my ocardium.Chick embryos, Hamburger-Hamilton (H-H) stage 8 to hatch, and 1 day old postnatal chicks were used in this study. For immunofluorescence study, each embryo was fixed with 4% paraformaldehyde and embedded in Epon 812. De-epoxinized with sodium methoxide, semithin sections were stained with primary antibodies (rabbit anti-desmin antibody and anti-vimentin antibody)and secondary antibody (RITC conjugated goat-anti rabbit IgG).

scholarly journals IMMUNOFLUORESCENT LOCALIZATION OF TWO DEHYDROGENASES IN HONEYBEE FLIGHT MUSCLE

1972 ◽  
Vol 20 (4) ◽  
pp. 266-271 ◽  
Author(s):  
RONALD W. BROSEMER
Keyword(s):  
Energy Metabolism ◽  
Flight Muscle ◽  
Immunofluorescent Localization ◽  
Glycerol 3 Phosphate Dehydrogenase

Two enzymes involved in energy metabolism, glycerol-3-phosphate dehydrogenase and triosephosphate dehydrogenase, were localized in honeybee flight muscle with immunofluorescent techniques. Both enzymes were found at or near the Z-band cross-striations.

scholarly journals Spindle dynamics and cell cycle regulation of dynein in the budding yeast, Saccharomyces cerevisiae.

1995 ◽  
Vol 130 (3) ◽  
pp. 687-700 ◽  
Author(s):  
E Yeh ◽  
R V Skibbens ◽  
J W Cheng ◽  
E D Salmon ◽  
K Bloom
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Nuclear Translocation ◽  
Spindle Pole ◽  
Cell Cortex ◽  
Wild Type ◽  
Immunofluorescent Localization ◽  
Yeast Saccharomyces Cerevisiae ◽  
Spindle Dynamics ◽  
Spindle Elongation ◽  
Anaphase B

We have used time-lapse digital- and video-enhanced differential interference contrast (DE-DIC, VE-DIC) microscopy to study the role of dynein in spindle and nuclear dynamics in the yeast Saccharomyces cerevisiae. The real-time analysis reveals six stages in the spindle cycle. Anaphase B onset appears marked by a rapid phase of spindle elongation, simultaneous with nuclear migration into the daughter cell. The onset and kinetics of rapid spindle elongation are identical in wild type and dynein mutants. In the absence of dynein the nucleus does not migrate as close to the neck as in wild-type cells and initial spindle elongation is confined primarily to the mother cell. Rapid oscillations of the elongating spindle between the mother and bud are observed in wild-type cells, followed by a slower growth phase until the spindle reaches its maximal length. This stage is protracted in the dynein mutants and devoid of oscillatory motion. Thus dynein is required for rapid penetration of the nucleus into the bud and anaphase B spindle dynamics. Genetic analysis reveals that in the absence of a functional central spindle (ndcl), dynein is essential for chromosome movement into the bud. Immunofluorescent localization of dynein-beta-galactosidase fusion proteins reveals that dynein is associated with spindle pole bodies and the cell cortex: with spindle pole body localization dependent on intact microtubules. A kinetic analysis of nuclear movement also revealed that cytokinesis is delayed until nuclear translocation is completed, indicative of a surveillance pathway monitoring nuclear transit into the bud.

Immunofluorescent localization of the ubiquitin-activating enzyme, E1, to the nucleus and cytoskeleton

1993 ◽  
Vol 264 (1) ◽  
pp. C93-C102 ◽  
Author(s):  
J. S. Trausch ◽  
S. J. Grenfell ◽  
P. M. Handley-Gearhart ◽  
A. Ciechanover ◽  
A. L. Schwartz
Keyword(s):  
Cell Lines ◽  
Chinese Hamster ◽  
Eukaryotic Cell ◽  
Nuclear Staining ◽  
Immunofluorescent Localization ◽  
Acid Protein ◽  
Ubiquitin Conjugation ◽  
Double Label ◽  
Pleiotropic Functions ◽  
Amino Acid Protein

Ubiquitin, a 76-amino acid protein, is covalently attached to abnormal and short-lived proteins, thus marking them for ATP-dependent proteolysis in eukaryotic cells. Ubiquitin is found within the cytoplasm, nucleus, microvilli, autophagic vacuoles, and lysosomes. The ubiquitin-activating enzyme, E1, catalyzes the first step in ubiquitin conjugation. To date, very little is known about the subcellular distribution of this enzyme. We have utilized immunofluorescence and immunoblotting to examine the cellular distribution of E1 in several eukaryotic cell lines, including HeLa, smooth muscle A7r5, choriocarcinoma BeWo, Pt K1, and Chinese hamster ovary (CHO) E36. E1 was identified in both cytoplasmic and nuclear compartments in all cell lines examined. However, the relative abundance within these compartments differed markedly between the cell lines. Even within a single cell line, nuclear distribution was not uniform, and certain cells demonstrated an absence of nuclear staining. E1 resides predominantly within the nucleus in BeWo. In contrast, its distribution in CHO and Pt K1 cells is mainly cytoplasmic. Within the cytoplasm, three pools of E1 were identified by double-label immunofluorescence. The first of these colocalized with phalloidin, indicating association of E1 with actin filaments. A second cytoplasmic pool colocalized with tubulin and was predominantly perinuclear in its distribution. The third pool associated with intermediate filaments. This suggests that E1 is associated with all three components of the cytoskeleton. The distribution of E1 was unaltered in a mutant line of CHO E36 designated ts20, in which the E1 can be thermally inactivated. The variable distribution of E1 among cell lines, including its apparent cytoskeletal association, suggests pleiotropic functions of this enzyme and the ubiquitin-conjugating system.

scholarly journals Deletions into an NH2-terminal hydrophobic domain result in secretion of rotavirus VP7, a resident endoplasmic reticulum membrane glycoprotein.

1985 ◽  
Vol 101 (6) ◽  
pp. 2199-2209 ◽  
Author(s):  
M S Poruchynsky ◽  
C Tyndall ◽  
G W Both ◽  
F Sato ◽  
A R Bellamy ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Integral Membrane Protein ◽  
Endoplasmic Reticulum Membrane ◽  
Wild Type ◽  
Positive Signal ◽  
Complex Type ◽  
Immunofluorescent Localization ◽  
Hydrophobic Domain ◽  
Rotavirus A ◽  
Altered Protein

Rotavirus, a non-enveloped reovirus, buds into the rough endoplasmic reticulum and transiently acquires a membrane. The structural glycoprotein, VP7, a 38-kD integral membrane protein of the endoplasmic reticulum (ER), presumably transfers to virus in this process. The gene for VP7 potentially encodes a protein of 326 amino acids which has two tandem hydrophobic domains at the NH2-terminal, each preceded by an in-frame ATG codon. A series of deletion mutants constructed from a full-length cDNA clone of the Simian 11 rotavirus VP7 gene were expressed in COS 7 cells. Products from wild-type, and mutants which did not affect the second hydrophobic domain of VP7, were localized by immunofluorescence to elements of the ER only. However, deletions affecting the second hydrophobic domain (mutants 42-61, 43-61, 47-61) showed immunofluorescent localization of VP7 which coincided with that of wheat germ agglutinin, indicating transport to the Golgi apparatus. Immunoprecipitable wild-type protein, or an altered protein lacking the first hydrophobic sequence, remained intracellular and endo-beta-N-acetylglucosaminidase H sensitive. In contrast, products of mutants 42-61, 43-61, and 47-61 were transported from the ER, and secreted. Glycosylation of the secreted molecules was inhibited by tunicamycin, resistant to endo-beta-N-acetylglucosaminidase H digestion and therefore of the N-linked complex type. An unglycosylated version of VP7 was also secreted. We suggest that the second hydrophobic domain contributes to a positive signal for ER location and a membrane anchor function. Secretion of the mutant glycoprotein implies that transport can be constitutive with the destination being dictated by an overriding compartmentalization signal.

scholarly journals Immunofluorescent localization of Tamm-Horsfall mucoprotein in human kidney

1969 ◽  
Vol 22 (3) ◽  
pp. 334-339 ◽  
Author(s):  
J. K. McKenzie ◽  
E. G. McQueen
Keyword(s):  
Human Kidney ◽  
Immunofluorescent Localization
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