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C Gilly

Publications and source records attributed to C Gilly.

16 recordsLinked to original sources
PubMed1994

Silver-staining of proteins in polyacrylamide gels: a general overview.

On the basis of the physico-chemical principles underlying silver-staining of proteins, which are recalled in this paper, several methods of silver-staining of proteins after SDS electrophoresis in polyacrylamide gels or isoelectric focusing were tested. The most valuable protocols are presented in this report, including standard methods for unsupported gels and new methods devised for thin (0.5 mm) supported gels for SDS electrophoresis or isoelectric focusing and for staining of small peptides. Generally speaking, the most rapid methods were found to be less sensitive and less reproducible than more time-consuming ones. Among the long methods, those using silver-diammine complex gave the most uniform sensitivity. They require however special home-made gels and cannot be applied to several electrophoretic systems (e.g. systems using tricine or bicine as the trailing ion, or isoelectric focusing in immobilized pH gradients). For these reasons, protocols based on silver nitrate are of a more general use and might be favored. Future trends for silver-staining will also be discussed.

Electrophoresis, Gel, Two-Dimensional↗
PubMed1989

Histone H1(0) mapping using monoclonal antibodies.

Monoclonal antibodies (mAb) to ox liver histone H1 degree were produced and characterized. Two sets of mice were immunized either with pure H1(0) or with an H1(0)-yeast tRNA complex. Eleven hybridomas of various clonal origin were selected. Typing of the antibodies indicated that all but three IgM belonged to the IgG1 class and contained kappa light chains. Immunoblotting experiments using peptides derived from H1(0) or H5 treated by various proteolytic agents (trypsin, N-bromosuccinimide, cyanogen bromide, acetic acid), revealed that nine of the mAb reacted with the globular part of H1(0). More advanced characterization of the antigenic determinants allowed us to determine distinct regions within this globular part which are involved in the antigenic recognition. The peptopes could be subdivided into two groups. Three mAb bound to residues 24-27 and were specific for H1(0). Six mAb bound to residues 27-30 and were specific for H1(0) except one of them which strongly cross-reacted with H5 and GH5. Two mAb reacted with the entire histone H1(0) but failed to react with any of the peptides, suggesting that the corresponding epitope is a conformational antigenic determinant. In order to confirm the localization of the two distinct regions which are involved in the antigenic recognition, a synthetic decapeptide corresponding to the beginning of human H1(0) globular part (from residue 19 to residue 28) was synthesized. Inhibition experiments of the reaction between H1(0) and the various IgG1 mAb by increasing amounts of peptide-bovine serum albumin conjugates were then performed.

Amino Acid Sequence↗
PubMed1983

Assembly kinetics of replicating chromatin: isolation and characterization of prenucleosomal and nucleosomal DNA.

Replicating chromatin is known to be more sensitive to micrococcal nuclease than bulk chromatin. We have used this property and a fractionation procedure based on the specific release of replicating material under mild micrococcal nuclease digestion, in order to analyse both the kinetics of maturation of newly replicated DNA into nucleosomes and the structure of the replicating material. As other authors, we initially observed that repetitive unit of newly replicated chromatin was shorter than that of bulk chromatin, however this result appears to be due to sliding of nucleosomes along the chromatin fibers close to the replicating fork. Replicative chromatin was fractionated and analysed. A prenucleosomal peak was observed and preliminary characterized.

Animals↗
PubMed1977

[Not Available].

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History of Pharmacy↗
PubMed1976

[Action of colchicine and of spreading on the width of the chromatid fiber].

Chromatid fibers obtained from cells previously treated with colchicine were found more condensed, with a 22% greater fiber diameter than those from untreated cells. Chromatid fibers obtained by the Langmir trough technique showed a greater degree of elongation than fibers obtained by simple sedimentation.

Biometry↗
PubMed1975

The metaphase chromosome ultrastructure. II. Helical organization of the basic chromosome fiber as revealed by acute angle metal deposition.

The acute angle metal depositon technique, discloses a helical chromosome fiber substructure. The microdensitometric analysis of the fiber parameters shows that a fibril 152 A in diameter coils to form the basic chromosome fiber 295 A in diameter with a pitch of 247 A. These figures should be corrected because of the thickening due to metal deposition, the real dimensions would be closer to 112 A for the fibril diameter and 255 A for fiber diameter. The real pitch value is probably somewhat smaller than 247 A (approximatively 200 A) as the values were obtained by analysing slightly stretched fibers. These results support the helical model of the chromosomes fiber organization and show that, most probably, the metaphase chromosome fiber as seen in electron microscopy is the second order of helical packing.

Chromium↗