Proper peptide handling and solubilization stands out as the place to start of a productive bioassay project, and we feel this handling guideline will allow you to dissolve your peptides the right way. On buy peptides usa with each peptide delivery, you might also observe reconstitution conditions which we have used in the peptide purification process – this is for your reference only, you might dissolve your peptide in an alternative solvent according to your assay needs.
– Use just a small aliquot of peptide to test the dissolution method. As soon as happy, apply to the bigger aliquot as needed.
– In principle, solvent used should be the solvent which is going to facilitate or be compatible with the experiment of yours. But, we shall also bear in mind that there might be a difficult task sometimes to find an “ideal” solvent which is going to solubilize peptides, maintain the integrity of theirs and work with biological assays.
-For initial solvent used should be the right one. For example, for a very hydrophobic peptide, it’s much better to dissolve it in a small amount of organic solvent (such as acetonitrile) or DMSO before using the aqueous solution. Quite simply, adding organic solvent to a suspension of hydrophobic peptide in aqueous solution will not be going to help very much in dissolving.
– Peptide remedy might be unstable at temperatures even smaller compared to -20°C. As a result, a peptide solution once prepared should be used as quickly as possible.
What solvent(s) I can use to dissolve my peptides?
If it’s a short peptide which is 5aa or less, try sterile sterilized water first and it is likely to dissolve.
For other peptides, the overall cost of the peptide will help determine which initial solvent to use. Assign a worth of -1 to acidic residues which include Asp(D), Glu(E), and also the C-terminal free acid(COOH). Assign a value of +1 to basic residues that include Arg (R), Lys (K), His (H), and the N terminal free amine(NH2). Calculate the overall cost of the whole peptide.
1. If the overall charge of the peptide is positive (a basic peptide), try to dissolve the peptide in sterile sterilized water first. If moisture fails, add ~20 % acetic acid solution. If the peptide still doesn’t dissolve, put drops of TFA (< 50ul), or use 0.1%TFA/H2O to solubilize the peptide. Then dilute the peptide solution to the preferred concentration. 2. If the overall cost of the peptide is negative (an acidic peptide), attempt to dissolve the peptide in sterile sterilized water first. If the peptide persists as noticeable particles, sonication can be tried. If water fails, add NH4OH (<50ul) or perhaps 0.1%NH4OH drop-wise. Then dilute the peptide solution to the desired concentration. If the peptide contains Cys, do not use basic methods (NH4OH), but use DMF alternatively. 3. Peptide whose general charge is 0 (the peptide is neutral). It generally dissolves in natural solvents, for instance, acetonitrile, methanol, or isopropanol. If this doesn’t dissolve completely: a) For peptides that tend to aggregate (due to the hydrophobic interaction), the fact of denaturants , for example, 8M urea or perhaps 6M guanidine HCl, may be also required. b) For extremely hydrophobic peptides (containing more than 75 % hydrophobic residues), add DMSO drop-wise (use DMF as an alternative for Cys with peptides), and then dilute the perfect solution with water to the preferred concentration. Storage Guideline Most lyophilized peptides shall be healthy at room temperature for a minimum of a few weeks. For long-term storage, it is strongly suggested you save peptide in powder form at -20°C or perhaps lower, away from effective light, and under dry condition. Repeated freeze-thaw cycles should be stayed away from. The shelf life of peptide solutions is limited, especially for peptides with cysteine(C), methionine(M), tryptophan(W), asparginine(N), glutamine(Q), or even N terminal glutamic acid(E). For example, a Cys containing peptide is easily oxidised, especially in basic conditions; certain residues are not difficult to racemise, such as Proline. Avoid DMSO if the peptide has Met, Trp or Cys, as a result of sulfoxide or perhaps disulfide formation. Peptide stability becomes worse when in a fix, particularly at the bigger pH (pH> 8). We therefore highly recommend trying to keep answers within the range of pH 4 6. It is advised that peptides containing methionine, cysteine, or even tryptophan residues be stored in oxygen-free atmosphere to avoid oxidation. The presence of dithiothreitol (DTT) can be helpful in preventing oxidation.