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41.
Monoclonal antibodies were prepared against two species of Methanomicrobiaceae. Antibody 1A is specific for Methanospirillum hungatei strain JF1 and the determinant it recognizes is expressed on the surface of JF1 cells, where it is exposed and accessible to antibody. The determinant is found in a polypeptide (MW<12,000) in the sheath that covers the bacterial cell; it is not present in Methanospirillum hungatei strain GP1; and it is not expressed on the surface of whole cells of the other 24 methanogenic bacteria tested. It is therefore a marker of strain JF1, consequently, antibody 1A is potentially useful for tracking JF1 and fragments thereof in a variety of samples. Antibody 7A is specific for Methanogenium cariaci JR1c. It did not react with any other methanogen tested, not even with Mg. marisnigri or Ms. hungatei JF1, although these cross-react with Mg. cariaci if tested with polyclonal antisera. Therefore antibody 7A recognizes specifically a marker of Mg. cariaci JR1c.Abbreviations SIA
slide immunoenzymatic assay
- SDS-PAGE
sodium dodecylsulfate polyacrylamide gel electrophoresis 相似文献
42.
43.
Esmail K. Shubber David Jacobson-Kram Dr. Jerry R. Williams 《Cell biology and toxicology》1986,2(3):379-399
Seven antischistosomal drugs, two antimalarial drugs, and one antiamoebic drug were tested in all five Ames strains for induction of mutation, as well as for induction of cytotoxicity, inhibition of cellular progression, and the induction of sister chromatid exchanges in two cultured mammalian cell lines. We found that two agents shown to be negative in the Ames test were positive for sister chromatid exchange induction. Based on qualitative and quantitative evaluation, we find that all but three of the pharmaceuticals should be considered to be potential human carcinogens.Abbreviations AA
2-aminoanthracene
- 9AACC
9-aminoacridine
- AM
amoscanate
- BrdUU
bromodeoxyuridine
- CA
chloroquine diphosphate
- CHO
Chinese hamster ovary
- CQ
chloroquine
- DAPI
46-diamidino-2-phenylindole
- DHY
dehydroemetine
- DMSO
dimethyl sulfoxide
- EB
ethidium bromide
- FCS
fetal calf serum
- FN
4-fluoro-3-nitrophenyl azide
- HY
hycanthone
- ICP
inhibiting cell progression
- LU
lucanthone
- MEM
minimal essential medium
- 2NF
2-nitrofurantoin
- 4NPD
4-nitro-O-phenylenediamine
- NZ
niridazole
- OL
oltipraz
- OX
oxaminiquine
- PBS
phosphate buffered saline
- PQ
primaquine
- PZ
praziquantel
- SA
sodium azide
- SCE
sister chromatid exchange 相似文献
44.
Aerobic ferrisiderophore reductase assay and activity stain for native polyacrylamide gels 总被引:4,自引:0,他引:4
The reduction of ferric iron from microbial iron-binding compounds (siderophores) releases the iron from the siderophore so that it may be utilized by the microorganism. A method to detect aerobic ferrisiderophore reductase activity using ferrozine as a ferrous iron trap is shown to be applicable to cytoplasmic fractions from Rhodopseudomonas sphaeroides and four other different species of bacteria. The ferrisiderophore reductase uses reduced nicotinamide cofactors as reducing agents, and activity is stimulated by flavins. This assay has been adapted as a staining method to locate ferrisiderophore reductase activity in native polyacrylamide gels. 相似文献
45.
M Krieger 《Analytical biochemistry》1983,135(2):383-391
Pores formed in the membranes of animal cells by complexes of sterols and the polyene antibiotic amphotericin B can efficiently kill the cells. Thus, in the absence of exogenous sources of cholesterol, inhibitors of enzymes in the cholesterol biosynthetic pathway render cells resistant to amphotericin B. Preincubation of Chinese hamster ovary cells with compactin or 25-hydroxycholesterol, inhibitors of the synthesis of the key intermediate mevalonate, protected cells from amphotericin B killing and this protection was reversed by the addition of exogenous mevalonate. The ability of compactin to confer amphotericin B resistance on normal cells was abolished when cells were provided exogenous cholesterol by the receptor-mediated endocytosis of low density lipoprotein. Low density lipoprotein receptor-defective Chinese hamster ovary cells were not subject to this low density lipoprotein-dependent amphotericin B killing. Exogenous mevalonate did not prevent 4,4,10 beta-trimethyl-trans-decal-3 beta-ol, an inhibitor of mevalonate conversion to sterols, from protecting cells from amphotericin B. A simple two-step protocol in which cells are preincubated (15-24 h) with potential inhibitors and then treated (3-6 h) with amphotericin B was devised to provide a sensitive method for detecting direct (e.g., competitive) and regulatory inhibitors of cholesterol biosynthesis. This protocol may prove useful in detecting potential antihypercholesterolemia drugs and is currently being used to isolate mutants in receptor-mediated endocytosis. 相似文献
46.
A method is presented for the rapid enzymatic determination of acetate in spent bacterial culture supernatants. The assay is based on a previously published assay for acetate kinase [Bergmeyer et al. (1974) in Methods of Enzymatic Analysis (Bergmeyer, H. V., ed.), Vol. 1, pp. 425-426, Verlag Chemie-Academic Press, New York/London], and is sufficiently sensitive to detect acetate levels of 50 microM. The assay is cheaper than commercially available assays and is particularly useful for occasional use by laboratories not equipped for routine acetate analysis using gas chromatography. The application of the assay to the measurement of acetate in bacterial cultures is described, though it should also be applicable to other biological fluids and foodstuffs. 相似文献
47.
Analysis of retinal and 3-dehydroretinal in the retina by high-pressure liquid chromatography 总被引:3,自引:0,他引:3
A sensitive analytical method was developed in order to study the rhodopsin-porphyropsin system in the eye. Oximes of 11-cis-retinal, all-trans-retinal, 11-cis-3-dehydroretinal, and all-trans-3-dehydroretinal were determined quantitatively by high-pressure liquid chromatography. This method was applied to the analysis of retinal and 3-dehydroretinal in the retinas of bullfrog and goldfish. The results agreed with those obtained from the bleaching kinetics of visual pigment extracted with detergent. A reliable result is obtained if the tissue contains more than 5 pmol of retinal (or 3-dehydroretinal). The chromophore composition could be determined in the eye of a small freshwater prawn, Palaemon pancidence, using 50 pmol of 11-cis-retinal and no 3-dehydroretinal. 相似文献
48.
A method for the assay of dehydroascorbic acid using high-performance liquid chromatography with uv detection is described. The dehydroascorbic acid is separated from ascorbic acid and reduced with dithiothreitol, and is then quantitated as ascorbic acid following rechromatography. Since as little as 22 pmol can be detected, sensitivity is at least 40-fold greater than that of other currently available procedures. This method was used to measure the level of dehydroascorbic acid in normal and chronic lymphocytic leukemia lymphocytes. A significantly higher concentration of dehydroascorbic acid was found in leukemic (21.80 +/- 3.55 nmol/10(8) cells, mean +/- SE) than in normal lymphocytes (9.32 +/- 1.15 nmol/10(8) cells) (P less than 0.03). Analysis of extracts from normal B cell lymphocytes revealed comparable dehydroascorbic acid levels to unfractionated lymphocytes, indicating that the elevated level in chronic lymphocytic leukemia was not simply a reflection of the increased percentage of B lymphocytes in this disorder. These studies illustrate that the technique can be used to measure the dehydroascorbic acid content from sources where only scanty material is available or low levels are found. 相似文献
49.
A highly sensitive enzymatic assay for diadenosine 5′,5?-P1,P3-triphosphate (Ap3A) has been established on the basis of the coupled luminescence assay for diadenosine 5′,5?-P1,P3-tetraphosphate (A. Ogilvie (1981)Anal. Biochem.115, 302–307). Snake venom phosphodiesterase splits Ap3A into AMP plus ADP which can be measured in a luminescence reaction containing pyruvate kinase, phosphoenolpyruvate and luciferin-luciferase. The procedure is linear with Ap3A levels ranging from 0.1 to 2 pmol. The assay has been used to measure Ap3A in various eukaryotic cells after ion-exchange chromatography and high-performance liquid chromatography of acidic extracts of the cells. The level of diadenosine triphosphate was higher in all instances than the level of diadenosine tetraphosphate. When growing in the abdominal cavity of mice, Ehrlich ascites tumor cells contained high amounts of Ap3A (), allowing direct optical determination in the HPLC chromatography. The quantitative measurement of Ap3A with the luminescence assay gave identical results. Ap3A extracted from Ehrlich cells was also chromatographed with authentic nucleotide in two thin-layer systems providing additional proof for the existence of Ap3A in biological material. 相似文献
50.