Reconstitution
Notice
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What is Reconstitution?
Reconstitution is the process of dissolving a lyophilized (freeze-dried) peptide powder in an appropriate solvent to create a solution suitable for use in research experiments. Since most research peptides are supplied as lyophilized powders for optimal stability during storage and shipping, reconstitution is a necessary preparation step before any experimental application. Proper reconstitution technique is essential for maintaining peptide integrity, achieving accurate concentrations, and ensuring reliable experimental results.
Step-by-Step Reconstitution Process
- Allow the vial to reach room temperature before opening to prevent moisture condensation on the cold peptide powder, which can cause degradation.
- Calculate the required volume of solvent based on the desired final concentration using the peptide's molecular weight and the amount of material in the vial.
- Add the solvent slowly along the inside wall of the vial rather than directly onto the peptide cake to avoid splashing or foaming.
- Allow the peptide to dissolve naturally. If needed, gently swirl the vial or use brief, gentle sonication. Avoid vigorous vortexing, which can denature certain peptides.
- Verify complete dissolution by visually inspecting the solution for clarity. A clear solution indicates complete dissolution.
Choosing the Right Solvent
The choice of reconstitution solvent depends on the peptide's physicochemical properties. Sterile water or bacteriostatic water is appropriate for most hydrophilic peptides carrying charged residues. For hydrophobic peptides, a small volume of DMSO (typically 50-100 microliters) can be used as an initial solvent, followed by dilution with aqueous buffer to the desired concentration. Dilute acetic acid (0.1%) is suitable for basic peptides, while dilute ammonium hydroxide can be used for acidic peptides. Always consult the supplier's recommendations for specific reconstitution guidance.
Post-Reconstitution Storage
Once reconstituted, peptide solutions are significantly less stable than their lyophilized counterparts. It is strongly recommended to aliquot the reconstituted peptide into single-use volumes immediately after preparation, then store the aliquots at -20C or below. This practice avoids the damaging effects of repeated freeze-thaw cycles. Reconstituted peptide solutions stored under refrigeration (2-8C) should ideally be used within a few days to a few weeks, depending on the specific peptide's stability profile.