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Author Lim L., C.
Source CiteSeerX
Content type Text
File Format PDF
Subject Domain (in DDC) Computer science, information & general works ♦ Data processing & computer science
Subject Keyword Serotonin Receptor Knock-out Agent ♦ Proteomic Profiling ♦ Direct Autoradiography ♦ Liquid Scintillation Counting ♦ Sds-polyacrylamide Gel ♦ Drug Bound ♦ Serotonin Receptor ♦ Linear Relationship ♦ Western Blotting ♦ Protein Marker ♦ Protein Band ♦ Novel Drug ♦ One-dimensional Sds-page ♦ Coomassie Brilliant Blue ♦ Liquid Scintillation ♦ Relative Mobility ♦ Calibration Plot ♦ High Count ♦ Molecular Weight ♦ Mammalian Mouse Brain ♦ Molecular Mass ♦ Pvdf Membrane
Abstract The binding of the novel drug 3H-LBT 3004, labeled with tritium 3H, to serotonin (5-HT) receptors was analysed using one-dimensional SDS-PAGE. Proteins from the mammalian mouse brain were separated using a 1 mm 10 % SDS-polyacrylamide gel. Following electrophoresis, the protein bands were detected using Coomassie Brilliant Blue staining technique. Three lanes were used for liquid scintillation counting in 3H quantification. The other three lanes were used for Western Blotting onto PVDF membrane that was utilized in direct autoradiography. Results of the liquid scintillation counting produced two peaks of high counts per minute. A calibration plot was also done to confirm the linear relationship between relative mobilities of protein markers and their known molecular masses. It was utilized in the determination of molecular weights of the proteins that produced the two peaks. Direct autoradiography showed no indication of binding between drug and protein, though liquid scintillation counting concluded that the drug bound to proteins other than serotonin receptors but does not conclude that the drug does not bind to serotonin receptors. The failure of detection of binding via autoradiography was accounted for and suggestions for improvement of experiment were given.
Educational Role Student ♦ Teacher
Age Range above 22 year
Educational Use Research
Education Level UG and PG ♦ Career/Technical Study