* Detailed instructions for making gels in-house can be found in Sambrook et al.
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For standard denaturing and reducing conditions, mix sample 1:1 by volume with Laemmli 2x sample buffer.
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Heat to 100oC for 5 minutes or 70oC for 10 minutes.
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Load 20-50 µg sample per lane, along with suitable positive and negative controls.
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Run the gel and transfer to PVDF or nitrocellulose membrane according to the manufacturer’s instructions for the equipment and materials in use.
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Confirm protein transfer by staining for 1 minute with Ponceau S, followed by complete destain in distilled water.
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Mark position of molecular weight standards if they are unlabeled.