|des(1-3)IGF-1 is a naturally occurring, endogenous protein, as well as drug, and truncated analogue of insulin-like growth factor 1 (IGF-1)|
|Synonyms||Somatomedin C, IGF-I, IGFI, IGF1, IGF-IA, Mechano growth factor, MGF, Des(1-3), Des1-3, Des 1-3, Des (1-3), IGF-1 (4-70)|
|MeSH Entry Terms|
|Chemical and physical data|
|Molecular Weight||7371.48 g·mol−1|
|Sequence||TLCGAELVDA LQFVCGDRGF YFNKPTGYGS SSRRAPQTGI VDECCFRSCD LRRLEMYCAP LKPAKSA.|
|Purity||Greater than 97.0%|
|Enthalpy of Vaporization|
|Index of Refraction|
|Solubility||It is recommended to reconstitute the IGF-I des(1-3) in sterile 18M-cm H2O not less than 100µg/ml, which can then be further diluted to other aqueous solutions.|
|Polar Surface Area|
|DSSTox Substance ID|
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RECOMMENDATIONS FOR SOLUBILIZATION OF PEPTIDES
Start with an appropriate choice of solvent that is both compatible with your experimental procedures and that does not react with or degrade the peptide. The solvent should be dry and degassed. Since most peptide work is small-scale, it is most practical to buy small volumes of dry solvent, rather than attempting to dry it yourself. Determine whether the peptide is acidic, basic or neutral and proceed with solubilization using a small amount of the peptide. Acidic and basic peptides are more soluble at neutral pH than acidic pH.
Peptides are acidic when the manufacturing process is complete, due to the presence of trifluoroacetate (TFA) as the counter ion (a result of the cleavage or purification process). Check whether the peptide is supplied as a TFA salt before you start the purification process.
Short non-hydrophobic peptides (less than 5 amino acids) and peptides containing >25% non-clustered, charged residues and <25% hydrophobic residues will typically dissolve in aqueous solutions.
Simply adding water may dissolve basic peptides. If it does not, first try a drop (10-20 µl) of glacial acetic acid and sonicate or vortex. This may be increased up to 30% acetic acid by volume for problematic sequences. Addition of base can also promote oxidation of cysteines to the disulphide so deoxygenated buffers should always be used. For basic peptides with net charge of +1 or greater, an acidic solution will be needed.
Acidic peptides with net charge of -1 or greater should be dissolved in a small amount of basic solvent such as 0.1% ammonium hydroxide or ammonium bicarbonate and diluted to the required stock concentration with water. The exception is peptides containing Cys, as disulphide bonds may form at alkaline pH.
Neutral or hydrophobic peptides with a net charge of zero can sometimes be brought into solution by addition of base, but often a gel forms. Generally this gel will only respond to dilution with higher amounts of distilled, deionized water, along with sonication and vortexing.
Peptides that are >50% hydrophobic may be difficult to dissolve in water alone and should be dissolved in a small amount of organic solvent, for example acetonitrile, methanol, isopropanol, dimethyl suphoxide (DMSO), dimethylformamide (DMF). This should be added drop wise, followed by sonication and vortexing after every drop until the peptide dissolves. The drop wise addition of the organic solvent can also be used for peptides that do not respond to pH adjustment. Note that some cell culture based assays may not react well to DMSO, so a different solvent should be considered.
Peptides that are >75% hydrophobic are unlikely to dissolve in aqueous solution alone and may require solubilization in a stronger solvent such as TFA or formic acid and at high concentration. The peptide may precipitate out when aqueous buffer is added. These conditions may not be compatible with some cell culture based experiments.
Organic solvents at certain concentrations are incompatible with some biochemical assays. A small amount of DMSO should be compatible with most immunological assays, but avoid DMSO if the peptide contains Methionine, Cysteine or Tryptophan due to sulphoxide or disulphide formation. These peptides should be prepared using 1,2-ethanediol (EDT) or dithiothreitol (DTT) in order to prevent oxidation. Oxygen-free water or buffers, or DTT are recommended for solubilizaton.Note, if DMSO solvent is at all wet it will not freeze at -20°C, so freezing at -30°C is recommended. DMSO is hygroscopic and will adsorb water if subjected to repeated freeze-thaw cycles. To minimize these issues, buy small volumes of dry DMSO for solubilzation purposes.
Peptides that are prone to aggregation may require strong denaturants (e.g., 6M urea or guanidine hydrochloride), which may then be able to be diluted.
If peptides are to be used in cellular assays, start by making a concentrated stock solution e.g., 5-10 mM, or 5-10mg/ml. Dilute the peptides into physiological buffer for use, such that the original solvent is present at no more than 0.1% in the final working solution. Diluted peptide solutions may also be filter-sterilized (0.22mm) if they are to be added to sterile cultures.
Store lyophilized protein at -20 °C. Aliquot the product after reconstitution to avoid repeated freezing/thawing cycles.
Reconstituted protein can be stored at 4 °C for a limited period of time. The lyophilized protein remains stable until the expiry date when stored at -20 °C.
WARNING! LABORATORY RESEARCH USE ONLY.
For research/laboratory in vitro use
Only Laboratory Reagents
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We strongly recommend that you follow these guidelines to ensure that substances retain their quality and potency.
Storage of Reconstituted Peptides
ALWAYS allow peptides to come to room temperature before reconstituting with water. This will avoid most cases of peptides becoming cloudy.
Allow peptides to warm to room temperature before opening the vial to prevent condensation of atmospheric moisture onto the peptide, making it easier to handle and weigh. It is recommended to dissolve only the required amount into a buffer. The remainder of the stock should be stored dry at -20°C.
Once the peptides have been mixed or reconstituted into a liquid solution they must be refrigerated at a temperature of between 2-8c. If stored at room temperature they may begin to degrade after as little as 24 hours. Therefore we recommend storing them in the fridge as soon as possible after mixing. Stored in the fridge they will remain potent for around 8 weeks before beginning to degrade. Do not shake peptides aggressively as this can ruin them. Store peptides in sealed containers away from food.
Storage in buffer is not recommended. Peptides in solution are only stable for up to one week when stored at 4°C.
Storage of Lyophilised (freeze-dried) Peptides
The peptides we sell come in a lyophilised, freeze-dried form. Like this, the peptides will remain stable for around 30 days at room temperature. If there not going to be used within this time then they must be stored in a freezer at a temperature of around -20c where they will remain stable for approximately 48 months.
If peptides are to be used frequently, solubilize and aliquot, then store frozen at -20°C to avoid freeze-thaw cycles. Typically the peptide will remain stable for several months.
For short term storage (up to 2 months), freeze at -20°C; for long term storage -80°C is recommended.
Ideally, vials for peptide storage should be high-density, polypropylene tubes, airtight, well-sealed with a screw cap with an O-ring. Avoid storing or making up peptide solutions in polystyrene containers as they adsorb peptide material to their surface.
Peptides containing Cys, Trp or Met are susceptible to oxidation. It is advisable to blanket the peptide with argon or nitrogen when the vial is opened. Buffers used to dissolve these peptides should be degassed. These peptides have limited stability; long-term storage is not recommended.
Hygroscopic peptides containing several charged residues (D, E, K, R, H) may take up water when exposed to air. To prevent this, argon or nitrogen may be used to blanket the peptide when the vial is opened. If inert gases are unavailable, then storage in a desiccator is a viable alternative. Before use, bring samples to room temperature in a desiccator.
Please ensure that your freezer is not ‘frost free’ or ‘auto defrosting’ as constant freeze/thaw cycles will degrade the peptides over time.
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