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  • 1
    Electronic Resource
    Electronic Resource
    Expression of the Human Interleukin–2 Receptor Gamma Chain in Insect Cells Using a Baculovirus Expression Vector (1995)
    Oxford, UK : Blackwell Publishing Ltd
    Scandinavian journal of immunology 41 (1995), S. 0 
    Details
    ISSN: 1365-3083
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: The gene encoding the γ-chain of the human lnterleukin–2 receptor was expressed in lepidopteran insect cells using the baculovirus expression vector system. The corresponding gene was inserted under the polyhedrin promoter of the Autographa californica nuclear polyhedrosis virus and expressed in the Spodoptera frugiperda insect cell line Sf9 during viral infection. The recombinant receptor protein was identified by immunoblotting in cell lysates, prepared from insect cells infected with the recombinant virus. At 40 h post infection the corresponding protein was detected as two major bands with apparent molecular weights of 50–60 kDa using a rabbit anti–human IL–2R γ-receptor specific antiserum. Metabolic labelling with [35S]–methionine and SDS–PAGE analysis of the recombinant baculovirus infected insect cells verified the immunoblotting data. The expressed IL–2Rγ-protein could also be determined on the surface of infected insect cells by flow cytometer analysis.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1365-3083.1995.tb03576.x
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  • 2
    Electronic Resource
    Electronic Resource
    Dual modulation of calcium channel current via recombinant α2-adrenoceptors in pheochromocytoma (PC-12) cells (1997)
    Springer
    Pflügers Archiv 435 (1997), S. 280-285 
    Details
    ISSN: 1432-2013
    Keywords: Key words Calcium channels ; α2-Adrenoceptors ; G proteins ; Dexmedetomidine ; PC-12 cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  The ability of recombinant rat α2D-and α2B-adrenoceptors expressed in nerve-growth-factor-differentiated pheochromocytoma PC-12 cells to modulate Ca2+ currents, recorded by the whole-cell patch-clamp technique, has been studied. Ca2+ currents in different cells were either reversibly reduced or increased by dexmedetomidine, an α2-adrenergic agonist, in a concentration-dependent manner. Pertussis toxin pretreatment reduced the number of cells that showed an inhibitory response and reduced the magnitude of inhibition. In cells expressing the α2B-adrenoceptor, pertussis toxin increased the proportion of cells from which a stimulatory effect on Ca2+ currents could be recorded. The magnitude of the inhibitory responses was unaffected but the stimulatory responses were considerably reduced by the dihydropyridine Ca2+ channel blocker nifedipine (5 μM). All effects of dexmedetomidine were reversible upon wash-out and inhibited by the antagonist rauwolscine. The results support the idea that modulation of voltage-dependent Ca2+ channels in transfected PC-12 cells is mediated by activation of recombinant α2D- and α2B-adrenoceptors. This receptor activation predominantly causes inhibition of dihydropyridine-insensitive Ca2+ channels via pertussis-toxin-sensitive G proteins. Additionally receptor activation can also lead to stimulation of dihydropyridine-sensitive Ca2+ channels via pertussis-toxin-insensitive mechanisms.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004240050513
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    Paper (German National Licenses)
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  • 3
    Electronic Resource
    Electronic Resource
    Endogenous γ-l-glutamylglutamate is a partial agonist at the N-methyl-d-aspartate receptors in cultured cerebellar granule cells (1995)
    Springer
    Neurochemical research 20 (1995), S. 1471-1476 
    Details
    ISSN: 1573-6903
    Keywords: Cerebellar neurons ; N-methyl-d-aspartate receptors ; γ-l-glutamylglutamate ; intracellular Ca2+ depolarization ; Mg2+ dependency
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract γ-l-Glutamylglutamate (LGG), an endogenous constituent of the brain, reduced the glutamateevoked increase in intracellular Ca2+ in cultured cerebellar granule cells. The extent and properties of this inhibition were different at different Mg2+ concentrations. The intracellular Ca2+ response to NMDA was slightly enhanced by 0.1 mM LGG in normal (1.3 mM) Mg2+ medium, but in Mg2+-free medium LGG was stimulatory at low (0.1–1 μM) NMDA and inhibitory at high (0.1–1 mM) NMDA concentrations. In the absence of Mg2+, LGG alone increased cytosolic free Ca2+ and depolarized the cells. These effects were potentiated by glycine and blocked by extracellular Mg2+, 2-amino-5-phosphonopentanoate (APV), 7-chlorokynurenate, 3-amino-1-hydroxypyrrolidin-2-one (HA-966) and 5,7-dinitroquinoxaline-2,3-dione (MNQX). The results indicate that LGG is a partial NMDA agonist. On the other hand, the non-NMDA antagonists 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) and 6,7-dinitroquinoxaline-2,3-dione (DNQX) also inhibited the effects of LGG. This indicates an involvement of non-NMDA receptors in the actions of LGG. The consequent depolarization may also contribute to the activation of NMDA receptor-governed ionophores.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00970596
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    Paper (German National Licenses)
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