Search results for "Immunodiffusion"
showing 9 items of 19 documents
Isolation of a high spin form of cytochrome P-450 induced in rat liver by 3-methylcholanthrene.
1983
Abstract A form of cytochrome P-450 (P-450 MC1) has been isolated from the livers of 3-methylcholanthrene-treated rats. The molecular weight is 54,500 and the heme iron is in the high spin configuration which clearly differenciates this form from the other major cytochrome induced by 3-methylcholanthrene (P-450 MC2). Whilst MC2 actively dealkylated 7-ethoxycoumarin and 7-ethoxyresorufin, MC1 was only active with 7-ethoxyresorufin. Ouchterlony immunodiffusion analysis and ELISA showed that anti MC1 and anti MC2 reacted with both MC1 and MC2 but preferentially with the homologous antigen. Both anti MC1 and MC2 cross-reacted strongly with microsomes from 3-methylcholanthrene, Aroclor 1254 and …
Studies on the Biosynthesis of Microsomal Membrane Proteins. Site of Synthesis and Mode of Insertion of Cytochrome b5, Cytochrome b5 Reductase, Cytoc…
1982
The site of synthesis and mechanism of insertion into membranes of several microsomal polypeptides was studied using translation system programmed in vitro with polysomes or with mRNA extracted from free and membrane-bound rat liver polysomes. Primary translation products of cytochrome b5, NADH: cytochrome b5 oxidoreductase, NADPH: cytochrome P-450 oxidoreductase and epoxide hydrolase were isolated by specific immunoprecipitation and compared with the mature proteins. The following observations were made: 1 While cytochrome b5 and NADH: cytochrome b5 oxidoreductase are synthesized in free polysomes, NADPH: cytochrome P-450 oxidoreductase and epoxide hydrolase are made in membrane-bound poly…
Arthus type inflammation with rat immunoglobulins.
1971
Etude comparative des proprietes biologiques des anticorps IgM, IgG1 et IgG2 du rat. Les resultats montrent que le pouvoir agglutinant et lytique des anticorps IgM est respectivement 37 et 100 fois superieur a celui des deux classes d'anticorps IgG. Par contre, en ce qui concerne le phenomene d'Arthus, les anticorps IgM sont moins actifs que les anticorps IgG, si les rapports sont exprimes en poids. Si l'on calcule les rapports par nombre de molecules, les anticorps IgM sont aussi, dans ce cas, plus actifs que les IgG.
Endogenous role of epoxide-hydratase. Development of a steroid epoxide-hydratase assay and properties of the enzyme.
1979
A highly sensitive and rapid radiometric assay for the determination of specific epoxide hydratase activity with a steroid epoxide (16α, 17α-epoxy-1,3,5(10)-estratrien-3-ol, ‘estroxide’) has been developed. The unreacted substrate was separated from the product 1,3,5(10)-estratrien-3,16β,17α-triol by extraction into light petroleum. The product was then extracted into ethyl acetate and measured by scintillation spectrometry. Radiochromatography established that after subtraction of the blank the entire radioactivity measured in the ethyl acetate phase resulted from the product 1,3,5(10)-estratrien-3,16,17-triol, whilst high performance liquid chromatography with the four possible isomers of…
Use of monoclonal and polyclonal antibodies as structural and topographical probes for hepatic epoxide hydrolase
1983
AbstractMonoclonal antibodies have been prepared against rat liver epoxide hydrolase (EH), some of which gave precipitation lines on immunodiffusion against pure EH suggesting the presence of repetitive structural domains on the enzyme. Using ELISA, with polyclonal antibodies to rat and rabbit liver EH, reactivity and therefore structural similarities between EH of all species tested, including human, were observed. This was in contrast to immunodiffusion results demonstrating the limitations of the latter technique. Using monoclonal antibodies in ELISA, greatest structural similarity was between rat, mouse, and Syrian hamster EH and relatively little between rat and human. Two of the antib…
The major isozyme of rat cardiac glutathione transferases. Its correspondence to hepatic transferase X.
1986
1. A major isozyme of rat heart glutathione transferase was purified to homogeneity by Sephadex G-200 gel filtration, ammonium sulfate precipitation, CM-cellulose chromatography and affinity chromatography on S-hexylglutathione-linked Sepharose 6B. 2. The purified isozyme was a dimer with an apparent relative molecular mass of 50000 composed of two Yb-size subunits (Mr= 26 500). The isozyme is immunologically related to rat liver glutathione transferase X and 3–3, especially closely to transferase X, and no immunological cross-reactivity with subunits 1 and 2 of hepatic glutathione transferases was observed. The isoelectric point (pI = 6.9) of the isozyme was identical with and the substrat…
Sugar specific cellular lectins of Phallusia mamillata hemocytes: Purification, characterization and evidence for cell surface localization
1989
Cellular lectins (CLs) of Phallusia mamillata were demonstrated in protein preparations obtained by salt fractionation from hemocytes sonicated in a suitable medium. Since the lectins from the precipitated fraction bind sugars containing D-galactosyl groups, they were purified by affinity chromatography on Sepharose. SDS-PAGE under reducing conditions showed that CLs are formed of two components of apparent MWs approximately 36,900 and 35,090 and thus differ from serum lectins (SLs) (MW about 62,200). The "shrinkage" observed when SLs were examined under nonreducing conditions suggest the presence of intrachain disulphide bonds which can affect the molecular structure of the SLs. CL-SL diff…
Immunonephelometric determination of the apolipoprotein A-II.
1989
A fully mechanized immunonephelometric method is described for the rapid and specific determination of apolipoprotein A-II in serum. The method utilizes commercially available sheep antiserum against human apolipoprotein A-II. Nephelometry was performed with the Behring Nephelometer Analyzer (BNA). A single determination can be performed in 12 minutes, requiring 10 microliters sample volume. The measuring range is about 0.08 to 1.25 g/l apolipoprotein A-II. Precision is characterized by intra-assay coefficients of variation of 3.37%, 3.93% and 4.49% for apolipoprotein A-II concentrations of 1.22 g/l, 0.376 g/l and 0.185 g/l, and inter-assay coefficients of variation of 4.27% for an apolipop…
Comperative studies with Culex pipiens egg rafts. Immunogenetic, electrophoretic and enzymatic analysis of unfertilized, compatible and incompatible …
1973
By applying immunologic, electrophoretic and enzymatic methods, extracts of different raft types of Culex pipiens were analysed. Rafts of the crosses Pa x Pa and Ha x Ha contained four common antigens, while unfertilized rafts of Pa and Ha (no antisera were prepared against them) and rafts of the crosses Og x Og, Og x Pa, and Pa x Og shared three common antigens with the remaining raft extracts. Disk-electrophoresis of raft extracts in acrylamide gel resulted in different electropherograms. Ten protein bands were common to all these raft types. The unfertilized rafts of Pa and Ha yielded three more protein bands, the crosses Pa x Ha and Ha x Pa one more, the crosses Og x Og and Pa x Og thre…