Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Dansyl Compounds”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 721 records · Page 40Linked to original sources
PubMed1992

Separation of monoclonal antibodies from antihapten antisera by two-dimensional affinity electrophoresis.

A high-resolution two-dimensional affinity electrophoresis (2D-AEP) method was developed, using capillary polyacrylamide gel (PAG) isoelectric focusing in the first and slab PAG affinity electrophoresis in the second direction. Using this method, anti-hapten antibodies were separated into a number of monoclonal antibody [immunoglobulin G (IgG)] families, each of which is composed of several IgG spots having an identical affinity to the hapten but different isoelectric points. 2D-AEP may offer a powerful tool for solving fundamental problems in immunochemistry such as antibody heterogeneity, its hapten binding specificity and antigen-dependent somatic mutation.

Antibodies, Monoclonal↗
PubMed1992

Rapid fluorimetric assay for the detection of the peptidyl alpha-amidating enzyme intermediate using high-performance liquid chromatography.

Peptidylglycine alpha-amidating enzyme catalyzes the conversion of glycine-extended peptides to their corresponding amidated peptides via a stable alpha-hydroxyglycine intermediate. Using a new rapid fluorimetric reversed-phase high-performance liquid chromatographic assay, we have demonstrated that the substrate and product of the amidation reaction, as well as both stereoisomers of the alpha-hydroxyglycine intermediate, can be separated and detected in quantities as low as 1 pmol. The method is highly reproducible and requires less than 11 min for separation and quantification.

Amino Acid Sequence↗
PubMed1994

Analysis of dansyl amino acids in feedstuffs and skin by micellar electrokinetic capillary chromatography.

Micellar electrokinetic capillary chromatography (MECC) using sodium dodecyl sulphate (SDS) and sodium cholate have been used for analyses of 30 dansylated (Dns) amino acids. The influences of sample preparation, Dns/amino acid ratio, sample solvent composition, and separation conditions including voltage, temperature, pH and buffer composition were investigated. Complete separations of acidic and neutral amino acids were obtained within 45 min in the SDS system. The efficiency expressed as number of theoretical plates for the applied capillary 0.52 m long were between 210,000 and 343,000, and the repeatability was very good with relative standard deviations on relative migration times between 0.09 and 0.70% and on relative normalised peak areas (RNPAs) between 0.85 and 3.41%. The linearity studies gave correlation coefficients between 0.9957 and 0.9993 for RNPAs against concentration. Detection limits were between 3 and 6 fmol or approximately 2 pg of each amino acid. Basic amino acids were separated in a MECC system using sodium cholate. Procedures and problems using Dns derivatisation for amino acids analysed by the MECC methods are described. Finally, examples of analyses of hydrolysates of real complex samples show, that this method can be applied to determine the amino acid composition of proteins in feedstuffs and skin.

Amino Acid Sequence↗
PubMed1994

Separation of 24 dansylamino acids by capillary electrophoresis with a non-ionic surfactant.

The separation of 24 dansylamino acids was investigated by capillary electrophoresis with an additive of micelles of a non-ionic surfactant, Tween 20. Although two pairs of peaks, norvaline and methionine derivatives, and didansyltyrosine and solvent (methanol), did not show good resolution, other dansylamino acids were well separated within 70 min using 100 mM Tween 20 and pH 2.40. The theoretical plate numbers calculated for dansylamino acids were 28,000-111,000 with a 19-cm capillary column.

Amino Acids↗
PubMed1996

Isocratic high-performance liquid chromatographic method for quantitative determination of lysine, histidine and tyrosine in foods.

A method for the quantitative determination of lysine, histidine and tyrosine in foods based on pre-column derivatization with 5-dimethylaminonaphthalene-1-sulfonyl chloride (DnsCl) and reversed-phase liquid chromatography has been developed. Derivatization conditions, including DnsCl concentration, time, temperature, and buffer solution were studied. To establish the reliability of the proposed liquid chromatographic (LC) method, the precision and accuracy of the analyses were evaluated using samples of casein and lysozyme.

Caseins↗
PubMed1977

Immunoadsorbents for the isolation of high-affinity anti-hapten antibodies in high yield.

Anti-hapten antibodies with affinities close to 10(8)/M can be isolated in high yield by elution under mild conditions from immunoadsorbents prepared by coupling haptens directly to an aminoethylamino (AEA) derivative of beaded agarose. In experiments with rabbit anti-2,4-dinitrophenyl (DNP) antibody with an average association constant (Ko) of 1.2 X 10(9)/M, elution of antibody bound to a Sepharose-AEA-DNP column produced a yield of 57%. With rabbit antibody to the 5-dimethylaminonaphthalene-1-sulfonyl (DANS) hapten with Ko=2.7 X 10(8)/M and a Sips heterogeneity index a=0.52, a 20% yield of the antibody found in the starting material by quantitative precipitin analysis was obtained by specific precipitation and dissolution of the precipitate with 0.18 M Na DANSulfonate; this antibody had Ko=6.6 X 10(8)/M and a=0.66. Purification of another aliquot of the same antibody preparation by immunoadsorption on DANS-AEA-Sepharose and elution with the same hapten concentration produced a yield of 75% of antibody with Ko=5.7 X 10(7)/M and a=0.29. In 3 production runs with the new method, apparent yields of 181, 101 and 94% (based on quantitative precipitin analysis of the starting material) of anti-DANS antibody were obtained. In 2 of these runs, the immunoadsorbent was further eluted with 0.1 M acetic acid and then with 0.1 M acetic acid in 10% dioxane; additonal yields of 9 and 4% were obtained with acetic acid and further yields of 15 and 11% with acetic acid/dioxane; the antibody eluted with acetic acid was damaged as shown by its failure to mediate passive immune lysis of DANS-coupled sheep erythrocytes, but the antibody eluted with acetic acid/dioxane was recovered intact.

Animals↗
PubMed1980

Agglutination microassay of hapten- or protein-modified liposomes using a multiple cell culture harvester.

A simple agglutination microassay is described for studying the presence of antigenic determinants on the surface of liposomal membranes. Antigen-sensitized radioactive liposomes, reacted with specific immune IgG in the wells of a microtest plate, were passed through a glass fiber filter in a multiple cell culture harvester. The amount of agglutinate trapped on the filter as determined by the radioactivity, was proportional to the amount of antigen in the liposomes and to the amount of specific immune IgG in the reaction mixture. The specificity of the agglutination was determined by inhibition with hapten and carrier antigens. Liposomes actively sensitized with hapten-conjugated lipid, and passively sensitized with protein, were used in the assay. A new method of preparing protein-bound liposomes is also described.

Agglutination Tests↗
PubMed1993

Epitope mapping of human immunoglobulin-specific murine monoclonal antibodies with domain-switched, deleted and point-mutated chimeric antibodies.

27 engineered chimeric antibodies possessing human gamma, epsilon, mu or alpha constant regions and V region specificity for nitrophenyl or dansyl were used to study the isotype specificity of 29 murine monoclonal antibodies (MAbs) specific for human immunoglobulins (IgG1-4, IgE, IgM, IgA or secretory piece). The isotype-restricted immunoreactivity observed with wild-type chimeric antibodies paralleled the pattern of each MAb's reactivity with purified human myeloma proteins. 16 mutant IgG anti-dansyl chimeric antibodies with genetically engineered domain switches, deletions or point-mutations were used as antigens to further characterize the epitopes recognized by the human IgG subclass-specific MAbs. The binding of three human IgG1-specific MAbs (HP6069, HP6070 and HP6091) was mapped to similar epitopes on the CH2 domain of human IgG1. Of the two anti-human IgG2 MAbs tested, HP6002 reacted with the CH2 of IgG2 while HP6014 bound to the CH1 domain. Both anti-human IgG3 MAbs (HP6047, HP6050) reacted with different regions of the IgG3 hinge. The anti-human IgG4 MAbs (HP6023, HP6025) bound to a similar epitope on the carboxyl terminus of CH2 or the CH3 of human IgG4. The three exclusion antibodies (HP6019, HP6030 and HP6058) bound to different epitopes in the CH2 domain of three of four IgG subclasses. The domain mapping was confirmed by competitive inhibition experiments. These results were used to select a group of IgG-reactive MAbs for construction of a poly-monoclonal anti-IgG capture and detection reagent that uniformly bound all four subclasses of human IgG. This study provides support for the use of engineered chimeric human chimeric antibodies as replacements for increasingly rare, purified human paraproteins in the specificity analysis of immunochemical reagents used in clinical and research laboratories for the detection and quantitation of human antibodies. Moreover, these studies demonstrate how the MAbs can serve as effective probes for examining conformational differences among the four human IgG subclasses.

Animals↗
PubMed1983

Interaction between rat alpha 1 fetoprotein and fluorescent derivatives of estrone in relation to the position and type of the fluorescent label.

A series of derivatives of estrone with fluorescent dyes (dansyl or coumarin) coupled at different positions on the steroid molecule, have been synthesized. These derivatives were tested for their quantum yields and their binding properties were determined with respect to rat alpha 1 fetoprotein. Derivatives at C-3 (estrone-3-hemisuccinate-dansyl-cadaverine and estrone-3-dansyl) compete with estrone for binding to the fetal protein; however derivates of estrone at C-6 (estrone-6-carboxymethyloxime-dansyl-cadaverine) and at C-17 (estrone-17-carboxymethyloxime-coumarine, estrone-17-carboxymethyloxime-dansyl-cadaverine and estrone-17-dansyl-hydrazine) compete poorly or not at all. The association constant of the radioactive derivative estrone-3-dansyl [3H] with rat alpha 1 fetoprotein was measured directly: the same number of high affinity binding sites (0.6) as that for estrone was found with an apparent association constant of 3.7 X 10(6) M-1. In addition to the high affinity binding sites, a low affinity class of binding sites was found which corresponds to the binding of the dansyl fraction of the fluorescent steroid derivative.

Animals↗