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1.
Klin Lab Diagn ; (4): 16-22, 35, 2002 Apr.
Article in Russian | MEDLINE | ID: mdl-12412503

ABSTRACT

Hydrocortisone, progesterone, testosterone, triiodothyronine, thyroxine, chorionic gonadotropin, prolactin, alpha-fetoprotein, luteinizing, follicle-stimulating, and thyrotropic hormones were measured in human sera and in Lyphochek Immunoassay Plus Control reference sera (Bio-Rad Laboratories, USA) using 4 commercial kits (Alkor Bio Inc. and Roche, automated analyzer Roche Cobas Core; DPC, automated analyzer Immulite; Bayer, automated analyzer ACS:180). Coordination and correlation between these kits was observed, the coordination decreasing in the series Alkor Bio/Bayer, Alkor Bio/Roche, and Alkor Bio/DPC.


Subject(s)
Immunoenzyme Techniques , Reagent Kits, Diagnostic , Hormones/blood , Humans , Linear Models , alpha-Fetoproteins/analysis
2.
Eur J Biochem ; 267(8): 2362-71, 2000 Apr.
Article in English | MEDLINE | ID: mdl-10759862

ABSTRACT

An Alu-DNA repeat-binding protein with a molecular mass of 68 kDa (p68) is identified in the somatic human cell nucleoplasm. Gel mobility shift assay (GMSA), South-western blotting and affinity purification on DNA attached to the carrier were used in the identification. GMSA revealed multiple complexes with the exponential dependence of their relative mobility. A narrow binding site of the p68 was revealed using synthetic oligonucleotides. It is located between the A-box and B-box of the RNA polymerase III promoter and is identical to that reported for the Alu-binding protein from human spermatozoids. The same narrow binding site, the similarity of the isolation procedure from germ and somatic cells, and similar binding properties and molecular masses suggest homology of the two proteins. Antibodies raised against Alu-protein complexes led to hypershift of the complexes in GMSA and stained p68 in active fractions in human spermatozoids and in Alu-RNA-containing alpha-RNP particles. Immunofluorescence of a HeLa cell monolayer revealed an intranuclear dot pattern with the dots corresponding to euchromatin areas and some dots located at the cell periphery in the cytoplasm. alpha-RNP particles bound Alu-DNA in vitro and contained p68 as shown using the immunogold procedure. Alu-DNA binding activity was revealed in cytoplasm as well as in nucleoplasm. The possible nature of the main Alu-DNA binding protein and its involvement in the particle structure are discussed.


Subject(s)
Alu Elements/genetics , DNA-Binding Proteins/chemistry , Nuclear Proteins/chemistry , Ribonucleoproteins/chemistry , Spermatozoa , Base Sequence , Binding Sites , DNA-Binding Proteins/genetics , Electrophoresis, Polyacrylamide Gel , Fluorescent Antibody Technique , HeLa Cells , Humans , Immunohistochemistry , Microscopy, Immunoelectron , Molecular Sequence Data , Oligonucleotides/metabolism , Promoter Regions, Genetic , RNA Polymerase III/genetics , RNA-Binding Proteins/chemistry , Sequence Alignment
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