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Practical guide

Reconstitution guide

Reconstitution is the process in which a lyophilized (freeze-dried) peptide is mixed with bacteriostatic water (BAC water) to obtain a liquid form ready for laboratory work. It is an inseparable part of preparing research peptides and requires a sterile environment, correct technique and accurate calculation of the solvent volume for the intended concentration.

Quick checklist

5 reconstitution steps

The procedure is summarised in five steps; for the complete guide with an explanation of each phase, see the detailed article.

  1. 1
    Sterile workspace preparation. Wipe the working surface with 70% isopropyl alcohol. Tools: lyophilized vial, BAC water, sterile syringe with needle, isopropyl alcohol swab.
  2. 2
    Stopper disinfection. Wipe the rubber stoppers of both vials (peptide + BAC water) with the swab and let dry for 10 seconds — wet alcohol would be pushed inside with the needle.
  3. 3
    Drawing BAC water. Using a sterile syringe, draw the calculated volume of BAC water. Without air bubbles.
  4. 4
    Adding to the peptide vial. Insert the needle at an oblique ~45° angle and let the liquid run down the wall of the vial, not directly onto the peptide pellet. This prevents foaming and denaturation.
  5. 5
    Dissolution. Rotate the vial gently between your fingers (do not shake). The lyophilizate dissolves within 30–60 seconds. After dissolution store in the refrigerator at 2–8 °C.
Watch out for

Most common mistakes

  • Shaking instead of rotating. Mechanical shaking can denature peptide structure.
  • Adding BAC water too quickly. Liquid hitting the pellet directly causes foaming and degradation of part of the peptide.
  • Using sterile water (0.9% NaCl) instead of BAC water. Sterile water has no preservative effect; a peptide in it lasts only 24 hours compared with approximately 28 days in BAC water.
  • Storing in the freezer after reconstitution. Repeated freezing and thawing degrades the peptide. After reconstitution it belongs in the refrigerator at 2–8 °C.
  • A sealed lyophilized vial stored at room temperature. Even the lyophilizate loses stability — it belongs in the refrigerator at 2–8 °C (short term) or freezer at −20 °C (long-term storage).
Before and after reconstitution

Lagerung

State Optimal temperature Stability duration
Lyophilizate (sealed vial) −20 °C long-term / 2–8 °C short-term 24–36 months at −20 °C
After reconstitution with BAC water 2–8 °C, protect from light ~28 days (peptide dependent)
After reconstitution with sterile water (no BAC) 2–8 °C 24 hours

Note: actual stability duration varies by peptide. Always consult the technical documentation of the specific batch.

Calculation

BAC water volume

For the prepared concentration a simple relation applies:

BAC water volume (mL) = peptide amount (mg) ÷ desired concentration (mg/mL)

Example: 5 mg of peptide dissolved in 2 mL of BAC water gives a concentration of 2.5 mg/mL.

For quick error-free calculation use the Peptide calculator — an interactive tool that computes the solvent volume and per-dose amount based on your experimental protocol.

Related content

Continue reading

FAQ

Häufig gestellte Fragen

What is bacteriostatic water?

BAC water is sterile water with 0.9% benzyl alcohol as a preservative. It prevents microbial growth in the reconstituted peptide and extends stability to approximately 28 days.

How long does a peptide last after reconstitution?

In BAC water at 2–8 °C a peptide is typically stable for 28 days. The exact duration depends on the peptide and is stated in the batch technical documentation.

What if I see flakes or cloudiness in the vial after reconstitution?

Discard from further use. Cloudiness indicates aggregation or microbial contamination. Record the batch number and contact the supplier.

Can I freeze a reconstituted peptide?

Not recommended. Repeated freeze-thaw cycles reduce peptide activity. If long-term storage is necessary, aliquot into smaller volumes and freeze once.

What if I do not have BAC water?

Sterile water (0.9% NaCl) is an alternative but stability drops to approximately 24 hours. For longer experimental protocols BAC water is the standard.

Disclaimer. This guide describes a standard laboratory procedure for working with lyophilized peptides in a research-only context. Molequa®® products are intended exclusively for laboratory and scientific research. They are not intended for human or veterinary use, diagnosis, treatment or consumption.
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