Anti-Biotin Antibodies Offer Superior Organelle-Specific Labelling of Mitochondria Over Avidin or Streptavidin

2008 ◽  
pp. 157-170
Author(s):  
Elisabeth D. Coene ◽  
Michael K. Shaw ◽  
David J. Vaux
Keyword(s):  
Specific Labelling

Scanning luminescence x-ray microscopy: Progress toward selective staining using microspheres

1994 ◽  
Vol 52 ◽  
pp. 74-75
Author(s):  
C. Jacobsen ◽  
J. Fu ◽  
S. Mayer ◽  
Y. Wang ◽  
S. Williams
Keyword(s):  
Visible Light ◽  
Active Sites ◽  
Light Emission ◽  
Chinese Hamster ◽  
Polystyrene Spheres ◽  
Good Resistance ◽  
X Ray ◽  
Selective Staining ◽  
Visible Light Emission ◽  
Specific Labelling

In scanning luminescence x-ray microscopy (SLXM), a high resolution x-ray probe is used to excite visible light emission (see Figs. 1 and 2). The technique has been developed with a goal of localizing dye-tagged biochemically active sites and structures at 50 nm resolution in thick, hydrated biological specimens. Following our initial efforts, Moronne et al. have begun to develop probes based on biotinylated terbium; we report here our progress towards using microspheres for tagging.Our initial experiments with microspheres were based on commercially-available carboxyl latex spheres which emitted ~ 5 visible light photons per x-ray absorbed, and which showed good resistance to bleaching under x-ray irradiation. Other work (such as that by Guo et al.) has shown that such spheres can be used for a variety of specific labelling applications. Our first efforts have been aimed at labelling ƒ actin in Chinese hamster ovarian (CHO) cells. By using a detergent/fixative protocol to load spheres into cells with permeabilized membranes and preserved morphology, we have succeeded in using commercial dye-loaded, spreptavidin-coated 0.03μm polystyrene spheres linked to biotin phalloidon to label f actin (see Fig. 3).

scholarly journals Identification of the protein responsible for pyruvate transport into rat liver and heart mitochondria by specific labelling with [3H]N-phenylmaleimide

1981 ◽  
Vol 196 (2) ◽  
pp. 471-479 ◽  
Author(s):  
A P Thomas ◽  
A P Halestrap
Keyword(s):  
Membrane Protein ◽  
Sodium Dodecyl Sulphate ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Mitochondrial Protein ◽  
Sodium Dodecyl ◽  
Liver Mitochondria ◽  
Heart Mitochondria ◽  
Approximate Amount ◽  
Specific Labelling

1. N-Phenylmaleimide irreversibly inhibits pyruvate transport into rat heart and liver mitochondria to a much greater extent than does N-ethylmaleimide, iodoacetate or bromopyruvate. alpha-Cyanocinnamate protects the pyruvate transporter from attack by this thiol-blocking reagent. 2. In both heart and liver mitochondria alpha-cyanocinnamate diminishes labelling by [3H]N-phenylmaleimide of a membrane protein of subunit mol.wt. 15000 on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. 3. Exposure of mitochondrial to unlabelled N-phenylmaleimide in the presence of alpha-cyanocinnamate, followed by removal of alpha-cyanocinnamate and exposure to [3H]N-phenylmaleimide, produced specific labelling of the same protein. 4. Both labelling and kinetic experiments with inhibitors gave values for the approximate amount of carrier present in liver and heart mitochondria of 100 and 450 pmol/mg of mitochondrial protein respectively. 5. The turnover numbers for net pyruvate transport and pyruvate exchange at 0 degrees C were 6 and 200 min-1 respectively.

The RNA content of the nucleolus and nucleolus-like inclusions in the anther of Lilium estimated by an improved RNase-gold labelling method

1989 ◽  
Vol 94 (4) ◽  
pp. 675-683
Author(s):  
S. SATO ◽  
C. WILLSON ◽  
H. G. DICKINSON
Keyword(s):  
Electric Discharge ◽  
Condensed Chromatin ◽  
Tetrad Stage ◽  
Dense Bodies ◽  
Rna Content ◽  
Predominant Type ◽  
Tapetal Cells ◽  
Gold Labelling ◽  
Labelling Method ◽  
Specific Labelling

Using RNase-gold labelling a comparative estimation has been made of the RNA content of the nucleoli and nucleolus-like inclusions in anther cells of Lilium henryi Thunb. Pretreatment with glow electric discharge prior to application of the RNase-gold labelling remarkably lowered the level of nonspecific adsorption and allowed specific labelling of RNA-containing structures. In the tapetal cells, the nucleoli, the perichromatin material and the dense bodies labelled heavily, but both the interchromatin region and the condensed chromatin did not bind the label. The nucleolus-like inclusions, which were found in the nucleoplasm of meiotic cells at the tetrad stage, and were termed ‘nuclear nucleolus-like bodies (NLBs)’ in the present study, also showed a high response compared with both the loosened and the condensed chromatin; this labelling was some 60% of that observed over the nucleoli of somatic cells. Another type of nuclear NLB was differentiated from the predominant type of NLB by a conspicuous electron opacity, reduced size, and strong labelling with the RNasegold complex, suggesting that some nuclear NLBs may be highly condensed. The labelling over the cytoplasmic nucleoloids (nucleolus-like inclusions found in the cytoplasm) was only 50 % of that over the nuclear NLBs, although both are similar in ultrastructural texture.

Specific Labelling of Phagosome-Derived Vesicles in Macrophages with a Membrane Dye Delivered with Microfabricated Microparticles

2021 ◽  
Author(s):  
Wenhao Cheng ◽  
Sundol Kim ◽  
Sandra Zivkovic ◽  
Hoyong Chung ◽  
Yi Ren ◽  
...  
Keyword(s):  
Specific Labelling

Selective reduction of a carboxyl group with diborane. Synthesis of specifically carbon-14 labelled D,L-2-Amino-4-(2-aminoethoxy)butanoic acid

1978 ◽  
Vol 56 (22) ◽  
pp. 2853-2855 ◽  
Author(s):  
Yu-Ying Liu ◽  
Edna Thom ◽  
Arnold A. Liebman
Keyword(s):  
Thionyl Chloride ◽  
Title Compound ◽  
Selective Reduction ◽  
Carboxyl Group ◽  
Butanoic Acid ◽  
Carbon 14 ◽  
Sodium Ethyl ◽  
Work Up ◽  
Specific Labelling

Specific labelling of the title compound was achieved through intermediate 3a, methyl 2-(2-phthalimidoethoxy)acetate, derived from methyl bromoacetate-2-14C. Conversion to the corresponding acid (4) was followed with selective diborane reduction which provided 2-(2-phthalimidoethoxy)ethanol-2-14C (5). Treatment with thionyl chloride yielded the chloride 6 which is identical with the intermediate used in the nonlabelled synthesis and reaction with sodium ethyl phthalimidomalonate provided the title compound after hydrolytic work-up.

Measurement in sheep of the quantity and composition of rumen digesta and of the fractional outflow rates of digesta constituents

1980 ◽  
Vol 31 (6) ◽  
pp. 1129 ◽  
Author(s):  
GJ Faichney
Keyword(s):  
Particulate Matter ◽  
Retention Time ◽  
Microbial Population ◽  
Quantitative Prediction ◽  
Representative Sampling ◽  
Mean Retention Time ◽  
Outflow Rate ◽  
Few Data ◽  
Specific Labelling

Knowledge of the fractional outflow rates of rumen digesta constituents is required for the quantitative prediction of rumen function. However, there are few data available for these parameters because of the problems associated with specific labelling of digesta constituents and with the need to ensure that samples are truly representative of rumen digesta. Methods are described here by which two markers can be used together to overcome the problem of representative sampling of the rumen, so eliminating the need for specific labelling, with 'ideal' markers, of digesta constituents. Thus, from the measurement of the amount and composition of true digesta in the rumen and the amount and composition of true digesta flowing from the abomasum, values for the fractional outflow rates (and its reciprocal, mean retention time) of rumen digesta constituents can be calculated. Appropriate application of these methods will allow the derivation of functions to predict fractional outflow rates. The results presented show that the fractional outflow rate of an adsorbable marker, 103Ru-P, was about half that of water but was about twice that of particulate matter; it was also greater than the net value for microbes. On the assumption that microbial material leaving the rumen is drawn from a free-floating microbial population and a population associated with particulate matter, it was calculated that 0.29 � SE 0.07 (range 0.09-0.49) of the rumen microbial population could have been free-floating.

Site-Specific Labelling of Native Mammalian Proteins for Single-Molecule FRET Measurements

ChemBioChem ◽  
2018 ◽  
Vol 19 (8) ◽  
pp. 780-783 ◽  
Author(s):  
Alexander Gust ◽  
Leonhard Jakob ◽  
Daniela M. Zeitler ◽  
Astrid Bruckmann ◽  
Kevin Kramm ◽  
...  
Keyword(s):  
Single Molecule ◽  
Site Specific ◽  
Single Molecule Fret ◽  
Mammalian Proteins ◽  
Specific Labelling

Polyacrolein microspheres as a new tool in cell biology

1982 ◽  
Vol 56 (1) ◽  
pp. 157-175
Author(s):  
S. Margel ◽  
U. Beitler ◽  
M. Ofarim
Keyword(s):  
Cell Biology ◽  
Polymerization Process ◽  
Amino Groups ◽  
Ph Values ◽  
Human Red Blood Cells ◽  
Primary Amino ◽  
Aldehyde Groups ◽  
Potential Use ◽  
Specific Labelling ◽  
Reactive Aldehyde

Polyacrolein (PA) microspheres in sizes ranging from 0.04 micron to 40 microns were synthesized. Magnetic and fluorescent PA microspheres were formed by carrying out the polymerization process in the presence of appropriate ferrofluidic or fluorochromic compounds, respectively. The microspheres carry reactive aldehyde groups, through which various ligands, containing primary amino groups, were covalently bound at physiological pH values. The potential use of these microspheres was demonstrated by the specific labelling of fresh human red blood cells (RBC) and by the separation of human RBC from turkey RBC by means of a magnetic field. PA microspheres were also bound covalently to the anti-allergic drug disodium chromoglycate (DSCG) and the conjugate was used for the labelling of rat basophilic leukaemia cells.

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