How to Reconstitute

Review Solubility Data:

• The solubility information provided by the peptide manufacturer or derived from the amino acid sequence should be consulted.

• Hydrophilic peptides typically dissolve readily in water, whereas hydrophobic peptides may require organic solvents. See which to use at the bottom of this page.

Prepare the Diluent:

• Use only sterile, research-grade diluents to ensure purity.

• Filter the solvent if necessary to remove potential particulates.

Calculate Required Volume:

• Determine the concentration needed for research applications (e.g., 1 mg/mL).

• Calculate the volume of solvent required using the formula:
  

• Use the formula:

  $$V = \frac{M}{C}$$

  Where:

  • V = Volume of solvent needed (mL)
  • M = Mass of peptide (mg)
  • C = Desired concentration (mg/mL)

  Example: If you have 5 mg of peptide and want a final concentration of 1 mg/mL, the calculation would be:

  $$V = \frac{5 \text{ mg}}{1 \text{ mg/mL}} = 5 \text{ mL}$$

  This means 5 mL of solvent is required to achieve the target concentration.

  If a higher concentration is needed, reduce the volume of solvent accordingly. Always use sterile, research-grade diluents for reconstitution.

Add Solvent Gradually:

• Introduce the solvent to the peptide vial slowly to avoid foaming or bubbling, which can denature the peptide.

• Gently swirl the vial to mix. Vigorous shaking should be avoided as it may affect peptide structure.

Adjust pH (If Necessary):

• For peptides requiring a specific pH for solubility, adjust the pH of the solution using sterile acid (e.g., hydrochloric acid) or base (e.g., sodium hydroxide).

• Monitor the pH with a calibrated pH meter.

Inspect the Solution:

• Ensure complete dissolution of the peptide. If particulates remain, additional solvent may be required.

• For peptides insoluble in water, a co-solvent like DMSO can be used in minimal quantities before dilution.

Aliquot and Store:

• Divide the reconstituted solution into smaller aliquots to minimize freeze-thaw cycles.

• Store aliquots at −20°C (-4°F) or below. Label with concentration, solvent used, and reconstitution date.