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At least 91 records · Page 5Linked to original sources
PubMed1983

Advantages of preelectrophoretic conjugation of polypeptides with fluorescent dyes.

A rapid simple procedure is described for the conjugation of proteins, glycoproteins, and peptides with the fluorescent dye fluorescein isothiocyanate during the time required to polymerize a polyacrylamide gel. Such conjugation does not perturb the electrophoretic mobility of the polypeptides in detergent containing gels. The location of polypeptide . dye conjugate is evident by inspection immediately upon removal of a gel from an electrophoresis cabinet avoiding the time required for postelectrophoretic staining and destaining procedures. The sensitivity of detection of polypeptide . fluorescein conjugates is at least equivalent to that obtained using Coomassie blue.

Animals↗
PubMed2000

Rapid staining of proteins on polyacrylamide gels and nitrocellulose membranes using a mixture of fluorescent dyes.

The present work describes a novel, fluorescence-based method for staining proteins on SDS-PAGE and membrane(s). In this method, proteins are stained using a mixed-dye (sulfo-rhodamine B and 1-anilino-8-naphthalene sulfonic acid (NH(4)(+))) solution. The mixed-dye staining protocol can detect proteins up to a concentration of 15 ng. This method is generally applicable to all proteins and is more sensitive than the conventional Coomassie blue method. The staining method is rapid and efficient. Staining-destaining of proteins using the mixed-dye protocol takes less than half an hour. Another interesting feature of the staining protocol described here is the applicability to the staining of proteins on nitrocellulose membranes.

Anilino Naphthalenesulfonates↗
PubMed1996

Assessment of the acrosome reaction in Bufo arenarum spermatozoa by immunostaining: comparison with other methods.

The acrosome reaction in Bufo arenarum spermatozoa has been visualised only by electron microscopy. Different staining procedures for spermatozoa are described in the present study. By light microscopy the acrosomal status cannot be determined. In some reacted sperm, stained with Coomassie blue, a filament that could be the "perforatorium' was observed, as seen by electron microscopy. Several fluorescent lectins were used, but only FITC-PNA binds to acrosomal proteins. However, the fluorescence observed was weak. Indirect immunofluorescence, with antibodies raised against acrosomal matrix, showed different staining patterns between acrosome-reacted and intact spermatozoa. The technique is specific for evaluating acrosomal status in a large population of spermatozoa. Moreover, immunostaining, in contrast with lectin staining, can be carried out in the presence of glycoconjugates such as oocyte extracellular matrix without interference.

Acrosome↗
PubMed2003

Aptamers switch on fluorescence of triphenylmethane dyes.

MG and SRB aptamers, which are short RNA sequences originally selected only for binding to malachite green or sulforhodamine B, can greatly enhance the fluorescence of normally nonfluorescent triphenylmethane dyes. MG aptamer enhances the quantum yields of malachite green (MG) and a novel rigidized derivative, indolinyl malachite green (IMG) by >2000-fold. SRB aptamer brightens patent blue V and VF by >90-fold. These enhancements are specific because MG aptamer has no effect on patent blue dyes and SRB aptamer has little or no effect on MG and IMG. Such sequence-specific fluorescence labeling of short RNA motifs is a first step toward genetically encodable fusion tags for imaging selected RNAs in vitro and in cells.

Fluorescence↗
PubMed2002

Analysis and quantitation of biotinylated apoB-containing lipoproteins with streptavidin-Cy3.

Non denaturing gradient gel electrophoresis (GGE) is commonly used to analyze the size distribution of lipoprotein particles. Its relatively low sensitivity and linear dynamic range limit use of GGE to quantify protein content of lipoproteins. We demonstrate a new high sensitivity method for analysis and quantitation of biotinylated apolipoprotein B (apoB)-containing lipoproteins using a fluorescent streptavidin-Cy3 conjugate and non covalent preelectrophoretic binding. Forty-four lipoprotein subfractions spanning the VLDL and LDL particle spectrum subfractions (11 each from four human subjects) were prepared by density gradient ultracentrifugation. An aliquot of each sample was biotinylated and GGE was performed. Gels also were stained for lipid with Oil Red O (32 samples) and for protein with Coomassie Brilliant Blue (30 samples). There was a significant relationship between the Cy3 fluorescent label area under the curve and the mass of apoB (P < 0.02-0.004) and total cholesterol (P < 0.03-0.004). Particle diameters of each absorbence/fluorescent peak were comparable between Oil-Red O and streptavidin-Cy3 treated biotinylated lipoproteins (+/-3.54 A, P = 0.3). Biotinylation and prestaining of lipoprotein particle with streptavidin-Cy3 provides a new fluorescence-based method for detection and quantitative analysis of lipoprotein subspecies by gradient gel electrophoresis.

Apolipoproteins B↗
PubMed1987

Nuclear events from fertilisation to the early cleavage stages in the domestic fowl (Gallus domesticus)

The changes in nuclear morphology from fertilisation to the 16-blastomere stage of development in the domestic fowl were examined in sections of the germinal disc stained with DAPI (4',6'-diamidino-2-phenylindole) or with the Feulgen technique. By one hour after the estimated time of ovulation, the second meiotic division of the ovum was completed and the spermatozoal heads (3-9) in the germinal disc had transformed into pronuclei. During the following two hours all the pronuclei enlarged, and two of them became juxtaposed. Mitotic figures and large pronuclei were observed at 4 hours. The zygote nucleus went through 3-4 division cycles in synchrony. The supernumerary male nuclei divided once, at most, and all showed evidence of degeneration by the 4-blastomere stage, at about five hours. Therefore, the nuclei destined to form the future chick embryo follow a pattern of behaviour during fertilisation and early cleavage similar to that of animals in general.

Animals↗