scholarly journals Trouble in VDAC field by putative inconsistencies of early antibody labeling studies

Channels ◽  
2009 ◽  
Vol 3 (6) ◽  
pp. 379-380 ◽  
Author(s):  
Friedrich P. Thinnes
Keyword(s):  
Antibody Labeling

Role of PKC isozymes in water transport in toad urinary bladder

1991 ◽  
Vol 49 ◽  
pp. 302-303
Author(s):  
A.J. Mia ◽  
L.X. Oakford ◽  
T. Yorio
Keyword(s):  
Urinary Bladder ◽  
Apical Membrane ◽  
Membrane Surface ◽  
Protein A ◽  
Cell Types ◽  
Leukemic Cells ◽  
Toad Urinary Bladder ◽  
Pkc Isozymes ◽  
Antibody Labeling

Protein kinase C (PKC) isozymes, when activated, are translocated to particulate membrane fractions for transport to the apical membrane surface in a variety of cell types. Evidence of PKC translocation was demonstrated in human megakaryoblastic leukemic cells, and in cardiac myocytes and fibroblasts, using FTTC immunofluorescent antibody labeling techniques. Recently, we reported immunogold localizations of PKC subtypes I and II in toad urinary bladder epithelia, following 60 min stimulation with Mezerein (MZ), a PKC activator, or antidiuretic hormone (ADH). Localization of isozyme subtypes I and n was carried out in separate grids using specific monoclonal antibodies with subsequent labeling with 20nm protein A-gold probes. Each PKC subtype was found to be distributed singularly and in discrete isolated patches in the cytosol as well as in the apical membrane domains. To determine if the PKC isozymes co-localized within the cell, a double immunogold labeling technique using single grids was utilized.

Use of Antibody to aid Visualization of Protein at the Termini of Bacteriophage Ø29 DNA

1980 ◽  
Vol 38 ◽  
pp. 540-541
Author(s):  
Dwight Anderson ◽  
Charlene Peterson ◽  
Gursaran Notani ◽  
Bernard Reilly
Keyword(s):  
Electron Microscopy ◽  
Bacillus Subtilis ◽  
Dna Synthesis ◽  
Restriction Endonuclease ◽  
Protein Complex ◽  
Protein Product ◽  
Viral Dna ◽  
Small Proteins ◽  
Antibody Labeling ◽  
Subtilis Bacteriophage

The protein product of cistron 3 of Bacillus subtilis bacteriophage Ø29 is essential for viral DNA synthesis and is covalently bound to the 5’-termini of the Ø29 DNA. When the DNA-protein complex is cleaved with a restriction endonuclease, the protein is bound to the two terminal fragments. The 28,000 dalton protein can be visualized by electron microscopy as a small dot and often is seen only when two ends are in apposition as in multimers or in glutaraldehyde-fixed aggregates. We sought to improve the visibility of these small proteins by use of antibody labeling.

Photoreception of Paramecium cilia: localization of photosensitivity and binding with anti-frog-rhodopsin IgG

1991 ◽  
Vol 99 (1) ◽  
pp. 67-72
Author(s):  
Y. Nakaoka ◽  
R. Tokioka ◽  
T. Shinozawa ◽  
J. Fujita ◽  
J. Usukura
Keyword(s):  
Membrane Potential ◽  
Polyclonal Antibody ◽  
The Other ◽  
Paramecium Bursaria ◽  
Potential Response ◽  
Light Stimulation ◽  
Intact Cells ◽  
Other Hand ◽  
Immunocytochemical Studies ◽  
Antibody Labeling

Paramecium bursaria is photosensitive and accumulates in a lighted area. The cells can be deciliated by a brief suspension in dilute ethanol. Both intact and deciliated cells showed depolarization in response to light stimulation by a step-increase from dark to above 0.7 mW cm-2 (550 nm). On the other hand, after a step-increase to below 0.4 mW cm-1, intact cells showed hyperpolarization, while the deciliated cells showed no change in membrane potential. This difference in membrane potential response between ciliated and deciliated cells suggests that both somatic and ciliary structures are photosensitive. In our search for the photoreceptive molecules, a polyclonal antibody induced in rabbits against frog rhodopsin was found to cross-react with a 63x10(3) Mr protein of P. bursaria, by immunoelectrophoresis. Immunocytochemical studies showed that the antibody labeling was localized on both the ciliary and the somatic membranes. These results raise the possibility that P. bursaria may contain a rhodopsin-like protein as a photoreceptor molecule.

scholarly journals Effect of lyophilization on HRP–antibody conjugation: an enhanced antibody labeling technology

2018 ◽  
Vol 11 (1) ◽  
Author(s):  
Saikant Regidi ◽  
Shilpa Ravindran ◽  
Ashitha L. Vijayan ◽  
Vani Maya ◽  
Lakshmi Sreedharan ◽  
...  
Keyword(s):  
Antibody Conjugation ◽  
Antibody Labeling

Optimization of antibody labeling with rhenium-188 using a prelabeled MAG3 chelate

2002 ◽  
Vol 248 (1-2) ◽  
pp. 173-182 ◽  
Author(s):  
José L Crudo ◽  
Martin M Edreira ◽  
Esteban R Obenaus ◽  
Marco Chinol ◽  
Giovanni Paganelli ◽  
...  
Keyword(s):  
Antibody Labeling

Rehydration of freeze substituted brain tissue for preembedding immuno-electron microscopy

2021 ◽  
Author(s):  
Linnaea E Ostroff ◽  
Janeth Perez-Garza ◽  
Emily Parrish ◽  
Zachary Deane
Keyword(s):  
Electron Microscopy ◽  
Full Potential ◽  
Serial Sections ◽  
Volume Reconstruction ◽  
Additional Advantage ◽  
Brain Circuits ◽  
Immuno Electron Microscopy ◽  
Neuronal Processes ◽  
Molecular Information ◽  
Antibody Labeling

Electron microscopy (EM) volume reconstruction is a powerful tool for investigating the fundamental structure of brain circuits, but the full potential of this technique is limited by the difficulty of integrating molecular information. High quality ultrastructural preservation is necessary for EM reconstruction, and intact, highly contrasted cell membranes are essential for following small neuronal processes through serial sections. Unfortunately, the antibody labeling methods used to identify most endogenous molecules result in compromised morphology, especially of membranes. Cryofixation can produce superior morphological preservation and has the additional advantage of allowing indefinite storage of valuable samples. We have developed a method based on cryofixation that allows sensitive immunolabeling of endogenous molecules, preserves excellent ultrastructure, and is compatible with high-contrast staining for serial EM reconstruction.

scholarly journals Fiber-optic immunosensor for measurement of myoglobin

1997 ◽  
Vol 43 (11) ◽  
pp. 2128-2136 ◽  
Author(s):  
C Michael Hanbury ◽  
W Greg Miller ◽  
Robert B Harris
Keyword(s):  
Fiber Optic ◽  
Response Times ◽  
Analytical Signal ◽  
Polyacrylamide Gel ◽  
Sensing Element ◽  
Core Diameter ◽  
Fluorescence Quench ◽  
Antibody Labeling ◽  
Gel Composition ◽  
Fluorescence Energy

Abstract A self-contained fiber-optic immunosensor was developed to measure the 16 500-Da protein myoglobin. The sensing element was constructed by entrapment of Cascade Blue-labeled antibody within polyacrylamide gel at the distal face of an optical fiber 300 μm in core diameter. The polyacrylamide gel composition was optimized to allow diffusion of myoglobin but to exclude hemoglobin and higher-molecular-mass proteins from the sensing area. The analytical signal was derived from fluorescence energy transfer between Cascade Blue and the heme group of myoglobin. Fluorescence quenching occurred when myoglobin bound to labeled antibody. The total amount of fluorescence quench was dependent on the antibody labeling conditions and the amount of antibody incorporated in the sensor gel matrix. Myoglobin concentrations >5 nmol/L (83 μg/L) were measurable with response times of 15 to 130 min limited by diffusion into the sensing element. This report demonstrates the technical feasibility for a self-contained immunosensor to measure a protein analyte.

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