> For the complete documentation index, see [llms.txt](https://docs.rplpeptides.com/llms.txt). Markdown versions of documentation pages are available by appending `.md` to page URLs; this page is available as [Markdown](https://docs.rplpeptides.com/docs/faq/reconstitution-faq.md).

# Reconstitution FAQ

## 1. What is the standard reconstitution protocol?

1. Remove lyophilized vial from -20°C storage
2. Equilibrate to room temperature (15–30 min, unopened)
3. Calculate required solvent volume for target concentration
4. Add solvent gently along the vial wall (not directly onto powder)
5. Cap and gently swirl or invert — do NOT vortex
6. Visually inspect for complete dissolution
7. Aliquot into single-use portions if required

## 2. Why should I avoid vortexing?

Vortexing introduces mechanical stress that can cause peptide aggregation and structural denaturation. Aggregated peptides may lose biological activity and produce artifacts in bioassays.

## 3. What is the recommended concentration range?

| Application                  |     Recommended Stock Concentration    |
| ---------------------------- | :------------------------------------: |
| Cell-based functional assays | 1–10 mM in DMSO or 1–5 mg/mL in buffer |
| In vivo studies              |   1–10 mg/mL in sterile PBS or saline  |
| Binding assays               |        0.1–1 mM in assay buffer        |
| Biophysical studies          |    0.5–5 mg/mL in appropriate buffer   |

## 4. How do I aliquot reconstituted peptide?

Protocol:

1. Prepare sterile, low-binding microcentrifuge tubes
2. Label each tube with product name, concentration, date, and batch number
3. Dispense calculated single-use volume (typically 10–50 µL per aliquot)
4. Snap-freeze in liquid nitrogen or dry ice/ethanol bath (optional, for long-term)
5. Store at -20°C

## 5. How many freeze-thaw cycles are acceptable?

Maximum 3 freeze-thaw cycles. Each cycle introduces ice crystal formation and concentration effects that can accelerate degradation. Single-use aliquots are strongly recommended.

## 6. How do I handle a peptide that aggregates after reconstitution?

If aggregation occurs:

* Dilute further — higher concentrations increase aggregation propensity
* Add a small amount of organic co-solvent (e.g., 0.1% DMSO or acetonitrile)
* Adjust pH away from the peptide's pI
* Consider adding 0.5 M arginine or 0.1% Tween-80 (check compatibility with downstream application)

***

*For reconstitution inquiries: <technical@rplpeptides.com>*


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