![]() Its suitability for detecting erythropoiesis-stimulating agents was examined and found to be suitable for anti-doping evaluation in sports events and for quantification of dystrophin levels in skeletal muscle of healthy controls and individuals with Becker and Duchenne muscular dystrophy. ![]() ProteinSimple Western and traditional western blot were compared for analyzing lipid-rich adipose tissue samples and were found to highly correlative. The system required less sample, antibody and time to run than conventional Western blot assay and was found to be highly sensitive, reproducible and quantitative over a large dynamic range. Western blotting and detection can be conducted in an integrated instrument through nano-volume capillaries (ProteinSimple). centrifuge at 12000 rpm at 4☌ for 10-15 min and collect supernatant for use.incubate the cell suspension on ice with shaking for 30 min.resuspend cell pellet with 1 ml prechilled RIPA buffer/10 7 cells.centrifuge at 2000 rpm at 4☌ for 5 min.remove the media and resuspend the pellet with 1 ml prechilled 1x PBS and transfer to 1.5 ml microtubes.transfer cells to prechilled 1.5 ml microtubes or 15 ml tubes and centrifuge at 2000 rpm at 4☌ for 5 min.(optional) remove 100 ul aliquot for cell counting.total protein should be stored at -20☌ until needed.centrifuge at 12,000 rpm at 4☌ for 10-15 min and collect supernatant for use.(optional) homogenize or sonicate thoroughly.scrape the cells completely and transfer to prechilled 1.5 ml microtubes on ice.add prechilled 400 ul-1 ml 1X RIPA buffer/100 mm dish.remove the supernatant and wash with 1X PBS to remove residual media.1x Transfer Buffer: 3 g Tris, 14.4 g Glycine and 200 ml methanol, add ddH 2O to 1L.The buffer should be either freshly prepared or prepared, aliquoted, and frozen for future use. ![]()
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