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  • 1
    ISSN: 1432-2048
    Keywords: Lepidium ; Membrane protein ; Monoclonal antibody TOP 71 ; Plasma membrane ; Tonoplast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Highly purified tonoplast and plasma-membrane vesicles isolated from roots of Lepidium sativum L. (garden cress) were used as a starting material for generating a monoclonal antibody against plant tonoplast. Tonoplast vesicles were isolated by discontinuous-sucrose-gradient centrifugation followed by free-flow electrophoresis. The deglycosylated tonoplast fraction was used to generate monoclonal antibodies by immunization of Balb/c-mice and by fusion of their β-lymphocytes with the mouse cell line X 63 Ag 8.653. Using plasma membrane purified by two-phase partitioning and freeflow electrophoresis to define the negative signal in screening, and purified tonoplast to define the positive signal in screening, a monoclonal antibody (TOP 71) was obtained which recognized a tonoplast protein of 71 kDa by immunoblotting in cress-root membrane fractions. Two-dimensional gel electrophoresis, affinoblotting and binding to concanavalin A showed that the TOP 71-antigen was a glycosylated protein and had an isoelectric point (pI) of 4.5. The TOP 71-antigen was found in the different tissues of organs of several higher plants (Glycine max L., Curcurbita pepo L., Zea mays L.) where it did not cross-react with the purified plasma-membrane fractions of these plants. Additionally, TOP 71 recognized its antigen in microsomal fractions of two lower plants (Chara globularis Thuili., Matteucia struthiopteris Tod.).
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-2013
    Keywords: Renal Hemodynamics ; Renal Autoregulation ; Microspheres
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Isolated rat kidneys were perfused with either a modified Krebs-Henseleit solution containing a gelatine preparation (Haemaccel, 35 g/l) or with a suspension of washed bovine red blood cells (RBC). When perfusion pressure (PP) was varied repeatedly in the range between 30 and 210 mm Hg autoregulation of renal plasma flow (RPF) was almost complete in RBC perfused kindneys. Changes of PP by steps of 20 mm Hg at intervals of 5 min resulted in an incomplete autoregulation of RPF and glomerular filtration rate (GFR). Renin release (RR) was inversely related to PP in the range between 50 and 150 mm Hg, while perfusion at a pressure below or above that range had no further effect on RR. The most marked increase in RR was obtained, when PP was reduced from 90 to 70 mm Hg. After reduction of PP, an increase in RR was measurable within 1 min, and a maximum was reached after 5 min. In kidneys perfused with a cell-free medium at a PP of 45 mm Hg for up to 30 min, RR remained elevated for the entire period of pressure reduction. Injection of microspheres into the renal artery resulted in a prompt decrease of RPF, GFR and urinary sodium excretion, but the values returned towards control levels within 15 min; RR increased only transiently after a short initial fall.
    Type of Medium: Electronic Resource
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