How to Reconstitute Peptides?

When talking about peptide reconstitution, we first need to explain why we need to reconstitute them in the first place.

Most peptides come in so-called lyophilized form. We already talked about peptide forms when we talked about how to store and handle peptides properly, so we won’t go into too many details.

Basically, this is the most stable form for storing and transporting peptides. Lyophilization is a process of removing water from the material; what we’re left with is a peptide in powder form which we package into our vials. This powdered peptide needs to be reconstituted (mixed with a solvent) before it can be used in the lab.

Why Would You Even Buy Peptides That Need Reconstitution?

Lyophilization is not a process strictly reserved for peptides. Pharmaceutical companies use it for the majority of the chemicals they sell simply because it allows for the most stable form of transport.

But why should you, as the end consumer, care about the way a peptide is transported and in which form? Well, there are several reasons which we’ll mention below:

Cost – of course, it all comes down to the price of a certain peptide. Powder (lyophilized) form will be a much cheaper option, rather than buying a pre-mixed peptide. Of course, if you are buying small batches, the savings might be negligible, but if you’re a part of a big lab, and doing many different peptide experiments, the savings can add up to quite a large sum.
Storage – we mentioned the topic of storage many times before, but it’s important to emphasize it because this, too, will save you a lot of money in the long run since powder form is easy to store and use on a later date. Just make sure to avoid thawing and refreezing.
Dosing – when you’re dealing with a pre-mixed peptide, you’re pretty much stuck with what you buy, and lack much room for customization. When you mix peptides yourself, you can adjust the solvent dosage per your specific requirement. This is what give you an extra layer of flexibility.

Peptide Reconstitution

Though peptide reconstitution is not a complex process, it needs to follow certain protocols to ensure it gets done successfully. Some of the recommended steps for most peptide reconstitution are:

Getting everything ready – before you begin the reconstitution process, you need to prepare everything you’ll need – a vial with lyophilized peptide, solvent, syringe and other vials and tubes you might need along the way.
Preparing the solvent – we commonly recommend bacteriostatic water to be used as a solvent, but you can use another one; just make sure it’s of high purity.
Calculations – after you have everything prepared, you need to calculate the peptide concentration you are looking to achieve. This calculator will help you in this process – Peptide Shop’s Peptide Calculator.
Reconstitution – once you have your concentration down, you need to slowly add the solvent to the vial with the lyophilized peptide. How much solvent you add to the vial, will directly depend on the concentration you are looking to achieve (and the previous step you took).
Mixing and shaking – once you’ve mixed the peptide with the solvent, you should avoid any vigorous shaking as this may cause denaturation of the peptide. Use only gentle swirling motions and keep on repeating those until the peptide fully dissolves. Gentle swirling is also good since the peptide is in a powder form and may become attached to the walls of the vial. This swirling will “pick up” all the peptide residue and make sure you’ve used every last gram of it.
Inspection – once the mixing is done, you need to visually inspect your peptide vial and check for any irregularities in color or visible particles. If you still see some peptide particles, you can heat the peptide gently. You can also use a process called sonication. Sonication is a process in which sound waves are used to agitate the particles in any given solution.
Usage – once fully reconstituted and dissolved, peptide is ready to be used for your desired lab experiment (since we only sell peptides for laboratory use).
Storage – we have an entire article on proper peptide storage which you should read through. Another thing (also mentioned in this article), is the importance of measuring out your peptides and splitting them into separate vials for long term storage. This way, you’ll only be reconstituting the exact amount of peptide you need and avoid freezing again.