For Science… From Science

Solubility Tips

Reconstituting a Peptide

  • Bring frozen or refrigerated peptides to room temperature in a desiccated chamber to avoid water absorption.
  • Always begin by reconstituting a small amount of peptide before committing the entire lot.
  • Use sterile water or sterile filtration.  If there are any Methionine (M), Cysteine (C), or Tryptophan (W) residues, use oxygen free solvents to prevent oxidation. 
  • Avoid reconstituting a peptide in a buffer, such as PBS.  Salts hinder solubility.
  • Choose the appropriate solvent.  Begin reconstituting at a concentration higher than your desired final working concentration.


Amino Acid Characteristics

Recommended Solvent

Hydrophilic residues (KRHDEN)


Hydrophobic residues (AVLIMFW)

Low solubility in aqueous solvents; are soluble in organic solvents (DMF, DMSO, TFA, Acetonitrile)


  • A solubilized peptide is completely clear. No flecks or cloudiness should be present.
  • If a peptide with more hydrophilic residues is still not completely reconstituted:
    • Adjust the pH of the solution according to the overall charge of the peptide
      • Count the possible POSITIVE charges (K, R, H, and free N-terminus)
      • Count the possible NEGATIVE charges (D, E, and free C-terminus)
      • Determine which is greater.
        • If positive charges are greater, add dilute acid dropwise to proteinate residues and maximize charge.
        • If negative charges are greater, add dilute base dropwise to deproteinate and maximize charge.
        • try sonication, gentle heat or an organic solvent, such as DMSO, Acetonitrile or DMF.

If your peptide is still not completely reconstituted, lyophilize the peptide and begin again.

Please contact NEP Technical Service for assistance, should you have difficulty with these recommendations.


"THANKS AGAIN for all the intellectual support that NEP has provided to us over the last year. My colleagues have been overwhelmingly impressed with the quality of service and level of thought that you have put into researching and thinking about our particular peptide and antibody production needs. The level of scholarship went well beyond what we expected from any peptide and antibody company, and was crucial in shaping our decisions. It has truly been a pleasure corresponding with you."
Cliff N. PhD - Lawrence Berkeley National Laboratory