A batch of samples has been added to the Loading Buffer, but has not yet been boiled for denaturation, and is currently stored at -20°C. It is expected to be used in a few weeks or even a month. How should it be preserved?
For protein samples that have been added to Loading Buffer but not yet boiled for denaturation, long-term storage requires balancing protein stability with compatibility for subsequent experiments. Specific recommendations are as follows:
I. Recommended Storage Conditions
Temperature:It is recommended to store at -80°C to minimize protein degradation and protease activity.
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Freezing method: Direct freezing in original tubes is acceptable without aliquoting. However, if sample volume is large, aliquoting into single-use portions is advised to avoid repeated freeze-thaw cycles.
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Precautions: Ensure that repeated freeze-thaw cycles are avoided during storage, as the presence of SDS in the Loading Buffer can cause SDS precipitation, sample non-uniformity, and abnormal gel electrophoresis.
II. Pre-use Treatment Recommendations
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Immediately boil after retrieval on the day of the experiment (95–100°C, 5 minutes);
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If SDS precipitation or layering is observed in the stored samples, gently vortex or briefly warm bath (37–50°C) before boiling to dissolve.
III. Additional Recommendations
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If the sample contains proteases (e.g., no Protease Inhibitor added) or is a less stable protein type (e.g., tissue lysate), consider adding protease inhibitors before freezing.
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