Peptide Reconstitution Guide

Proper reconstitution of lyophilized peptides is crucial for maintaining their activity and ensuring accurate results in your research. This guide provides general procedures and recommendations for reconstituting CHPeptides products.

IMPORTANT DISCLAIMER: The reconstitution guidelines provided below represent general practices in the research field and are provided for informational purposes only. CHPeptides does not guarantee any specific results and is not responsible for any damages, losses, or injuries that may result from following these guidelines. Each research environment is unique, and researchers should use their professional judgment when reconstituting and handling peptides. These guidelines do not constitute professional advice for specific research applications.

General Reconstitution Principles

Before beginning reconstitution:

1. Allow the peptide vial to reach room temperature before opening
2. Review the product-specific data sheet for any special instructions
3. Use appropriate personal protective equipment
4. Work in a clean environment to avoid contamination

Selecting the Appropriate Solvent

The choice of solvent depends on the peptide's characteristics:

Hydrophilic Peptides

• Primary Solvent: Sterile distilled water or buffer
• pH Consideration: Use buffer with pH 5-7 for optimal stability

Basic Peptides (containing Arg, Lys, His)

• Primary Solvent: Dilute acetic acid (0.1%)
• Follow-up: May be further diluted with water or buffer

Hydrophobic Peptides

• Initial Solvent: DMSO, DMF, or acetonitrile (10-20%)
• Secondary Dilution: Dilute with water or aqueous buffer
• Note: Final DMSO concentration should be compatible with your experimental system

Difficult Cases

• Approach: Start with a small amount of DMSO (≤10%)
• Follow-up: Dilute with aqueous buffer
• Alternative: Use 0.1% TFA followed by dilution with buffer

Step-by-Step Reconstitution Procedure

1. Calculate the volume needed for your desired concentration

   • Example: For 1mg peptide at 1mM, calculate based on molecular weight

2. Add solvent slowly

   • Add solvent down the side of the vial
   • Start with a small volume (50-70% of final volume)

3. Dissolve the peptide

   • Gently swirl or rotate the vial
   • Avoid vigorous shaking or vortexing
   • Allow sufficient time for complete dissolution

4. Address dissolution issues

   • For incomplete dissolution, try gentle warming (37°C water bath)
   • Brief sonication (1-2 minutes) may help
   • pH adjustment within a biocompatible range can improve solubility

5. Adjust to final volume

   • Add remaining solvent to reach desired concentration
   • Mix gently to ensure homogeneous solution

Peptide-Specific Considerations

Cysteine-Containing Peptides

• Consider adding a reducing agent (e.g., DTT or TCEP) to prevent disulfide formation
• Use degassed solvents when possible

Hydrophobic Peptides

• May require higher percentages of organic solvents initially
• Gradual dilution into aqueous systems to prevent precipitation

Aggregation-Prone Peptides

• Consider addition of 10-100mM ammonium bicarbonate
• Addition of 5-10% acetic acid may help certain peptides

Storage After Reconstitution

For reconstituted peptides:

1. Immediate Use: Use freshly prepared solutions when possible
2. Short-term Storage: 2-8°C for up to one week
3. Long-term Storage:
   • Prepare small aliquots
   • Store at -20°C or -80°C
   • Avoid repeated freeze-thaw cycles

Troubleshooting

Peptide Won't Dissolve

• Try an alternative solvent system
• Adjust pH within 2 units of the peptide's isoelectric point
• Consider gentle warming or brief sonication

Solution Appears Cloudy

• Indicates potential aggregation or precipitation
• Try more suitable solvent combination
• Filtration may be necessary (note this may reduce peptide concentration)

Peptide Activity Loss

• Check pH of solution (extremes can denature peptides)
• Minimize exposure to freeze-thaw cycles
• Consider presence of proteases in buffers

Documentation

Always record:

• Reconstitution date
• Solvent used
• Concentration
• Storage conditions
• Any observations about solubility or appearance

REMINDER: These reconstitution guidelines are provided for general research purposes only. CHPeptides makes no representations or warranties regarding the suitability of these guidelines for your specific research applications. Always use proper laboratory safety procedures when handling research materials.