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3) For large volume of supernatant, concentrate the proteins by ammonium sulphate precipitation, dialyze the dissolved protein solution against 1 PBS, and then apply the solution onto the GSH column

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Press the yellow cap firmly to release the powder into the ampoule 2

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Also, the release of Ca 2+ from the mitochondria can be mediated by extracellular ATP bound to microglial purinergic receptors