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| [1.] Iam/Fragment 047 01 - Diskussion Zuletzt bearbeitet: 2014-03-12 20:01:29 Graf Isolan | Fragment, Gesichtet, Iam, SMWFragment, Schutzlevel sysop, Sheikh 2006, Verschleierung |
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| Untersuchte Arbeit: Seite: 47, Zeilen: 1ff (complete) |
Quelle: Sheikh 2006 Seite(n): 51, 54, 56, Zeilen: 51:5-24; 54:14; 56:1-7.15-17 |
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| 3.3.3.g. Assay procedure
The whole procedure was performed at room temperature. All samples, standards and controls were brought to the RT and assayed in duplicates. To synchronize the reaction in each well, all reagents were pipette using a multi-channel pipette. 50 μl of assay diluent was added to each well. Standards, control and samples were added in a quantity of 50 μl per well. The components were mixed by gentle tapping the plate frame for 1 min. After that, the plate was covered with the adhesive strip provided and incubated for 2 h at room temperature. After the incubation period each well was aspirated and washed with 400 μl of wash buffer using a manifold dispenser, and procedure was repeated four times for a total of five washes. After washing, 100 μl of rat CXCL2 conjugate was added to each well. The plate was covered with a new adhesive strip and incubated for another 2 h at room temperature. The aspiration and washing procedure was performed as described above. Subsequently, 100 μl of substrate solution was added to each well to start enzymatic reaction and plate was incubated in the dark for 30 min at room temperature. To stop the enzymatic reaction, 100 μl of stop solution was added to each well, followed by determination of optical density of each well using a microplate reader (Dynatech Laboratories) set to dual wavelength mode (test filter –450 nm, reference filter –570 nm). The calculation of results was performed with a program (Dynatech MRX software, version 1.33) created in accordance to the manual instructions (Quantikine® immunoassay kit). 3.4 Methods in clinical chemistry 3.4.1 Transaminases Transminases (ALT and AST) are the most important representatives of a group of enzymes, the aminotransferases or transaminases, which catalyze the conversion of α-keto acids into amino acids by transfer of amino groups. As a liver specific enzyme AST is significantly elevated in hepatobiliary disease, increased ALT levels, however can occur in connection with damages of heart or skeletal muscle as well as of liver parenchyma. Serum level of transaminases (AP, ALT and AST) were determined by routine clinical laboratory test using diasys kit (diagnostic systems international Holzheim Germany) |
[Page 51]
3.3.1.VIII Assay procedure The whole procedure was performed at room temperature. All samples, standards and controls were brought to the RT and assayed in duplicates. To synchronize the reaction in each well, all reagents were pipette using a multi-channel pipette. 50 μl of assay diluent was added to each well. Standards, control and samples were added in a quantity of 50 μl per well. The components were mixed by gentle tapping the plate frame for 1 min. After that, the plate was covered with the adhesive strip provided and incubated for 2 h at room temperature. After the incubation period each well was aspirated and washed with 400 μl of wash buffer using a manifold dispenser, and procedure was repeated four times for a total of five washes. After washing, 100 μl of rat IL-6, IL-1β, TNF-α or IFN γ conjugate was added to each well. The plate was covered with a new adhesive strip and incubated for another 2 hours at room temperature. The aspiration and washing procedure was performed as described above. Subsequently, 100 μl of substrate solution was added to each well to start enzymatic reaction and plate was incubated in the dark for 30 min at room temperature. To stop the enzymatic reaction, 100 μl of stop solution was added to each well, followed by determination of optical density of each well using a microplate reader (Dynatech Laboratories) set to dual wavelength mode (test filter 450 nm, reference filter 570 nm). The calculation of results was performed with a program (Dynatech MRX software, version 1.33) created in accordance to the manual instructions (Quantikine® immunoassay kit). [Page 54] 3.4 Methods in clinical chemistry [Page 56] 3.4.2 Transaminases Transminases (ALT and AST) are the most important representatives of a group of enzymes, the aminotransferases or transaminases, which catalyze the conversion of α- keto acids into amino acids by transfer of amino groups. As a liver specific enzymes ALT is significantly elevated in hepatobiliary disease, increased AST levels however, can occur in connection with damages of heart or skeletal muscle as well as of liver parenchyma. [...] Serum level of transaminases were determined by routine clinical laboratory test using diasys kit (diagnostic systems international Holzheim Germany) |
But for minutiae (that adapt the existing text to Iam's research subject) identical, nevertheless not marked as a citation. |
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Letzte Bearbeitung dieser Seite: durch Benutzer:Graf Isolan, Zeitstempel: 20140312200213