One widely used method for protein separation via electrophoresis employs a discontinuous polyacrylamide gel as the support medium and utilizes sodium dodecyl sulfate (SDS) to denature the proteins. This method is commonly known as sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). To begin, you’ll need two gel casts, each comprising four gel plates, four gel plastics, and eight spacers. Assemble these components and secure them with the gel cast cover. Be cautious and mindful of the acrylamide kit, which should be retrieved from the refrigerator and handled in a fume hood due to potential health risks associated with excessive inhalation. The first step involves preparing the resolving gel mix, designed for the separation of proteins based on their molecular weight. In a beaker, combine the following ingredients: 38.80 mL of nano-pure water, 19.88 mL of acrylamide, 20.00 mL of resolving buffer, 760 µL of SDS, 560 µL of APS, and 44 µL of TEMED. Carefully agitate the solution and set your pipettor to 5 mL for transferring the prepared solution (the resolving gel mix) into the …

Here’s the protocol for preparing and running 30 percent SDS-PAGE gels: 1. Retrieve the SDS gels as these are the ones we’ll be using for this procedure. 2. Prepare your extracted samples by gathering the following items: a tray with crushed ice, pipettor, 96-well plate, extraction buffer, sample buffer, and the protein extracts (the samples that have been extracted). Thaw the samples and ensure they are fully defrosted. Use a vortex to thoroughly mix the samples. 3. Take 20 microtubes containing protein extracts and place them in a centrifuge. Spin the centrifuge for five minutes to separate any debris. 4. Label the 96-well plate accordingly, designating ten wells for rice and ten for maize samples. 5. With the pipettor set to 50 µL, add the sample buffer to each well. 6. Reset the pipettor to 35 µL and add the extraction buffer to the designated wells. 7. Reset the pipettor again to 15 µL and add the protein extracts to each well, where the sample and extract buffers have been placed. 8. Heat the 96-well …