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ANALYTICAL METHODS · PRACTICAL GUIDE

Diluting a Peptide Stock? Count the Final DMSO Percentage

Track how much DMSO enters the final mixture when you dilute a peptide stock.

NHD Technical Team3 teaching figuresSources checked: 2026-10-11

A small addition of a concentrated peptide stock also adds its solvent. If the stock contains DMSO, you need to track that liquid as well as the peptide. The two calculations answer different questions.

Count the solvent volume first

DMSO is a solvent sometimes used in research stock solutions. To calculate its final volume percentage, find how many microlitres of DMSO were added, then divide by the final mixture volume.

A stock made in pure DMSO contributes one microlitre of DMSO per microlitre transferred. A stock containing 20% DMSO contributes less.

Final DMSO fraction · 1
Figure 1. Both peptide and solvent travel with the stock aliquot.

An example with pure DMSO

Assume a 1 mM peptide stock in pure DMSO. Transfer 10 µL and make the final mixture up to 1,000 µL. The peptide concentration is 1,000 µM × 10 ÷ 1,000 = 10 µM.

The DMSO percentage is 10 ÷ 1,000 × 100% = 1% by volume. These numbers describe the same mixture, but one describes peptide concentration and the other solvent proportion.

10 µL DMSO ÷ 1,000 µL final volume × 100% = 1%

Final DMSO fraction · 2
Figure 2. In this example, 10 µL of pure DMSO in a final 1,000 µL gives 1% DMSO.

What changes if the stock contains 20% DMSO?

The same 10 µL aliquot now carries 10 × 0.20 = 2 µL of DMSO. In a final 1,000 µL mixture, that is 0.2% by volume.

If several reagents contain DMSO, add all their DMSO contributions before dividing by the final volume. Do not count only the peptide stock.

Why a solvent control matters

If wells receive different stock volumes, their DMSO percentages can differ too. A suitable solvent control helps you investigate the solvent contribution instead of attributing every change to the peptide.

Whether a particular percentage is acceptable depends on the research system. The 1% and 0.2% here are calculations, not recommended limits.

Final DMSO fraction · 3
Figure 3. Calculate solvent exposure alongside the peptide concentration.

Try an aliquot of a mixed-solvent stock

Transfer 5 µL of a stock containing 40% DMSO into a final 1,000 µL. What is the final DMSO percentage?

See the answer

The aliquot contains 5 × 0.40 = 2 µL of DMSO. The final percentage is 2 ÷ 1,000 × 100% = 0.2% by volume.

Write the stock solvent composition and final volume next to the calculation. A stock volume alone is not enough to reproduce it.

Check the solvent when planning LL-37 or SS-31 work

For LL-37 or SS-31, confirm which solvent is suitable for the supplied material. Then calculate what that solvent contributes to your planned final mixture. A product name does not establish a universal DMSO preparation method.

LL-37 / SS-31

Discuss material and document requirements with NHD · Continue with the analytical guide