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Measuring protein levels of receptors and enzymes involved in endocannabinoid metabolism is a significant step toward understanding the production, function, and regulation of these components of the endocannabinoid system. Western blotting is a common technique for detecting proteins from complex biological systems. With a focus on finding type 1 cannabinoid receptors, we present a general introduction to western blotting protein components of the endocannabinoid system using sodium dodecyl sulfate gel electrophoresis and nitrocellulose membranes in this chapter.
Source link: https://doi.org/10.1007/978-1-0716-2728-0_32
Duchenne muscular dystrophy is a degenerative muscular disease that affects approximately one in 5,000 males at birth. New advances in gene therapy and the recognition of dystrophin expression in other muscular dystrophies have opened new avenues for therapy. DMD and Becker muscular dystrophy are both common muscle dystrophies, requiring precise monitoring of dystrophin expression and determining the safety of new therapies for muscular dystrophies. Here, we describe the testing of a novel Western blot technique for the quantitation of mini-dystrophin protein in human skeletal muscle tissues, which is simple to implement in most laboratory settings. Step 1 involves protein extraction from skeletal muscle tissue lysates from control, DMD, or BMD biospecimens. Step 3 involves running gel electrophoresis with wild-type dystrophin from muscle tissue extracts along the mini-dystrophin STD curve and mini-DYS and protein normalization with housekeeping GAPDH.
Source link: https://doi.org/10.1208/s12248-022-00776-0
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