Reconstituting Lyophilised Proteins: A Practical Bench Protocol
A short, practical protocol for bringing lyophilised proteins back into solution without losing activity.
Before you open the vial
Spin the vial briefly in a microcentrifuge so that powder adhering to the cap is collected, and let the vial reach room temperature to avoid condensation on the powder.
Choosing the diluent
Use the buffer recommended on the CoA. If none is given, a neutral buffer such as PBS is usually safe; only add carrier protein (for example BSA) if your assay is compatible with it, since carriers can interfere with calibrators.
Adding the liquid and storing
Add the diluent down the wall of the vial, never directly onto the powder, and mix by gentle inversion rather than vortexing. Prepare single-use aliquots before freezing to avoid repeated freeze–thaw cycles, and record the lot number on every aliquot.
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