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At least 613 records · Page 34Linked to original sources
PubMed1986

A study of the messenger RNA encoding pyruvate kinase of Neurospora crassa.

In Neurospora crassa, there is a single pyruvate kinase (PK) consisting of four identical subunits of approximately 60k daltons. Northern and dot blot hybridization studies, using most of the yeast pyruvate kinase gene as a probe, suggest the presence of two distinct mRNA species for pyruvate kinase, separable on the basis of the length of their polyadenylated tails, by oligo(dT)cellulose chromatography. These messages are present in polysomes, immuno-precipitated by anti-PK antibodies, indicating probable translation in vivo. Fractions containing both messages were translated in vitro in the heterologous systems as well as in a homologous N. crassa lysate, the newly-synthesized PK being detected by immunoadsorption. Protection studies using S1-nuclease suggest no major structural differences in the 5'-untranslated and most of the coding regions of the two messages.

Animals↗
PubMed1985

High expression levels of ras p21 protein in normal mouse heart tissues.

We have investigated the levels of protein encoded by the ras oncogene in normal mouse tissues using an immunoblotting technique. We have found that heart from young or adult NIH or Balb C strains of mice contain high levels of ras protein as compared to lung, liver, spleen, kidney, brain and skeletal muscle tissues from the same animal. Our results indicate that cellular ras expression does not in every case correlate with cell proliferation.

Animals↗
PubMed1987

Analysis of epidermal proteins for DNA-binding activity.

A group of DNA-binding proteins from the soluble extract of newborn rat epidermis have been separated by chromatography using DNA-cellulose columns. The electrophoretogram of the DNA-binding proteins eluted from a single stranded DNA-cellulose column shows five major proteins of molecular weights ranging between 25K to 40K. Both the epidermal protein filaggrin and most keratins, except two high molecular weight keratins, do not show in vitro DNA-binding activity.

Animals↗
PubMed1986

Partial characterization of human complement factor H by protein and cDNA sequencing: homology with other complement and non-complement proteins.

Factor H, a control protein of the human complement system, is closely related in functional activity to two other complement control proteins, C4b-binding protein (C4bp) and complement receptor type 1 (CR1). C4bp is known to have an unusual primary structure consisting of eight homologous units each about 60 amino acids long. Such units also occur in the N-terminal regions of the complement proteins C2 and factor B, and in the non-complement serum glycoprotein beta 2I. Amino acid sequencing, and sequencing of a factor H cDNA clone, show that factor H also contains internal repeating units, and is homologous to the proteins listed above.

Amino Acid Sequence↗
PubMed1986

Immunological detection of cDNA clones encoding the uncoupling protein of brown adipose tissue: evidence for an antigenic determinant within the C-terminal eleven amino acids.

Poly(A)+RNA was isolated from brown adipose tissue of cold acclimated rats and a fraction enriched for uncoupling protein mRNA was used to generate a cDNA library in pBR 322. Immunological screening of 1,500 colonies with an affinity-purified antiserum against the uncoupling protein yielded five positive clones, pUCPrat1-5. Clone pUCPrat2 encoded the C-terminal 54 amino acids of rat uncoupling protein and exhibited 90% amino acid homology with the hamster protein. Clones pUCPrat3-5 encoded only the C-terminal 11 amino acids suggesting that an antigenic determinant lies within this sequence.

Adipose Tissue↗
PubMed1988

Distribution of protein kinase C immunoreactivity in rat retina.

A polyclonal antiserum to protein kinase C has been used to study the distribution of the enzyme antigenic sites in rat retina. The results indicate that the kinase is concentrated in photoreceptor outer segments as well as in the outer and inner plexiform layers. In identified components of retinal neuronal circuits, the kinase immunoreactivity is present in photoreceptor presynaptic terminals, in bipolar cell dendrites and axons, and probably in bipolar cell presynaptic terminals impinging on retinal ganglion cell dendrites. Thus, protein kinase C is positioned to play a role in specialized compartments of photoreceptor membrane and at both pre- and postsynaptic levels in the function of retinal neuronal circuits. Label in the nucleus is observed in retinal ganglion cells, but not bipolar or horizontal cells and probably not in amacrine cells. A role for protein kinase C in neuronal function at the level of the cell nucleus is therefore not likely to be universal, but to be determined by the particular properties of individual neuronal types.

Animals↗
PubMed1975

Excess electrical noise during current flow through porous membranes separating ionic solutions.

Spectral analysis of electrical noise from various artificial membrane systems suggests that excess noise of an f-n spectral form, where n is approximately unity, is not primarily a bulk phenomenon simply dependent on the number of charge carriers. Measurements from aqueous and nonaqueous electrolytic resistors, comprised of several different ionic species, show only flat power density spectra under applied currents, even at extreme dilutions. Excess noise of f-n form is observed under applied d-c current in single pore membranes, as previously reported, but is also seen in multipore and polymer mesh membranes. Calculations based on single pore membrane noise data are in significant variance with the bulk charge carrier model proposed by Hooge. These observations suggest that such excess noise occurs in conjunction with anisotropic constraints to ion flow.

Collodion↗
PubMed1983

The effect of anti-inflammatory and anti-rheumatic drugs on cell accumulation onto nitrocellulose discs implanted subcutaneously into mice.

Nitrocellulose discs were implanted subcutaneously into mice and cell accumulation measured 96 hours later. Cell accumulation was inhibited by the anti-inflammatory steroids hydrocortisone, dexamethasone and triamcinolone acetonide, but higher doses of the latter two compounds were required than are normally used in rat models of inflammation. Cell accumulation was not inhibited by the cyclo-oxygenase inhibitors naproxen, indomethacin, tolmetin or aspirin, nor by the dual cyclo-oxygenase and lipoxygenase inhibitors BW 755C and benoxaprofen. Anti-rheumatic drugs such as D-penicillamine, dapsone and colchicine and the metal chelator 1,10 phenanthroline were all inactive on cell accumulation in this model.

Animals↗
PubMed1987

Enzyme-linked immunofiltration assay for rapid serodiagnosis of syphilis.

A new rapid technique for detection of serum treponemal antibodies is described which is based on an enzyme-linked immunoassay using nitrocellulose as solid phase. With this technique antigen-antibody binding is accelerated by the filtration of the antibody solution through the antigen-coated nitrocellulose filter instead of its remaining over the solid phase for incubation. Test results are available in less than 15 min. Serum specimens from 255 syphilitics and 829 non-infected subjects were investigated. The sensitivity and specificity of the Treponema pallidum enzyme-linked immunofiltration assay were comparable to those of the Treponema pallidum haemagglutination assay and the fluorescent treponemal antibody absorption test.

Antibodies, Bacterial↗
PubMed1996

Identification of glycoproteins on nitrocellulose membranes and gels.

In this article we describe a procedure for the detection of glycoproteins on gels employing the periodic acid-Schiff's reagent. In addition, a number of staining protocols and direct binding ELISA, employing antibodies and lectins, are described for the identification and quantitation of glycoproteins after their immobilization by dot, slot, or Western blotting onto nitrocellulose membranes. We document, in detail, the conditions (i.e., the effect of solvent and detergents) for the immobilization of one specific family of O-linked glycoproteins, namely mucins. However, taking into account our suggestions, these procedures should be applicable to other types of glycoprotein.

Acrylic Resins↗
PubMed1986

Visualization of c-Ki-ras-2 oncogene sequences in human pancreas, a chemically induced transplantable carcinoma, and carcinomas of pancreas by in situ hybridization.

c-Ki-ras-2 sequences were visualized in paraffin-embedded sections from normal fetal and adult human pancreases, a chemically induced transplantable human pancreas carcinoma (PT-1) and three carcinomas of pancreas by in situ hybridization technique. A biotinylated 1-kilobase-pair (kb) EcoRI fragment of pHiHi3 DNA was used as probe and the oncogene was visualized as one or two large grains of reaction products produced by streptavidin-peroxidase complex and diaminobenzidine tetrachloride in more than 9% of normal pancreas nuclei. Its amplification in the chemically induced cell line was detected as one or more large grains in 72% of the nuclei and numerous cytoplasmic grains. The detection of oncogene in normal pancreases and its amplification in PT-1 cells was validated by Southern analysis of EcoRI digests of genomic DNA extracted from normal pancreases and PT-1 cell line. The oncogene was also demonstrated to be equally amplified in two adenocarcinomas and one undifferentiated carcinoma of human pancreas by in situ hybridization.

Adenocarcinoma↗
PubMedDate not supplied

Effect of nickel(II) on DNA-protein interactions.

Alterations in DNA-protein interactions (DPI) may play an important role in carcinogenesis. Although the mechanism of nickel carcinogenesis is unknown, nickel reportedly affects DPI. A microfiltration, nitrocellulose filter assay was utilized to study DPI in intact Chinese hamster ovary (CHO) cells and in isolated nuclei. Prior to exposure of CHO cells or isolated CHO cell nuclei, DNA and proteins were radiolabeled using 3H-thymidine and 35S-methionine, respectively. Nuclei were exposed to NiCl2 in 10 mM HEPES buffer (pH 6.8). CHO cells were exposed in either complete or a salts-glucose medium. Following exposure, nuclei or cells were incubated at 37 degrees C for 20 min in a high salt lysis solution; aliquots were loaded onto nitrocellulose filters and washed with a low salt solution. DNA (3H) retained on each filter was normalized to protein (35S) bound on the filter. Exposure of either whole cells or isolated nuclei to increasing, noncytotoxic concentrations of NiCl2 resulted in a dose dependent decrease in DPI. The effect of nickel on specific DNA-protein interactions was examined using a band shift assay and a cloned satellite DNA sequence. Nickel inhibited specific protein binding to the satellite DNA probe. The results of these two independent assays, which were conducted at physiological pH, indicate that NiCl2 inhibits specific DNA-protein interactions.

Animals↗
PubMed2001

Potentiometric phosphate-sensing system utilizing phosphate-binding protein.

A phosphate-detection system has been developed which uses phosphate-binding protein (PBP) from Escherichia coli. PBP was immobilized on a sheet of nitrocellulose membrane by cross-linking and the membrane potential of the immobilized PBP was measured. The response time of the system to phosphate was 5 min. The response was selective to phosphate among other anions. Under optimum conditions 0.1-1.5 mmol L(-1) phosphate can be determined with this system.

Biosensing Techniques↗