What will happen if the protein lysis buffer is placed on ice for 48 hours?
Leaving protein lysate on ice for 48 hours can lead to several situations, depending on the composition of the lysate.
1. Situations with relatively stable conditions (most commercial or freshly prepared lysates)
If it contains basic components such as buffer (e.g., Tris-HCl), detergents (e.g., NP-40, Triton X-100), and salts (e.g., NaCl):
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There is usually no significant change after being left on ice for 48 hours;
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The lysate can still be used normally, especially when operating in a cold room;
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It is recommended to gently mix or briefly centrifuge before use to avoid local precipitation or stratification.
2. Situations where stability might be affected
1. Contains reducing agents (e.g., DTT, β-mercaptoethanol)
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They are easily oxidized in solution, and their activity may significantly decrease after 48 hours;
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It is recommended to prepare fresh for immediate use, or store below -20°C to prevent inactivation.
2. Contains protease/phosphatase inhibitor cocktail
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Inhibitors may become partially inactive or degrade;
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It is recommended to add them immediately before use.
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