I have 10 mg of peptide synthesized from company in China and it came in lyophilized form. I need to prepare a stock solution (1mg/ml) and then have to use it in very small concentrations such as 64 µg/mL.
I am confused that if it won't get dissolved completely (homogenous mixture) then how I will be able to take out this small concentration? Usually, primers get dissolved in nuclease free water.
Please guide me.
Dear Zeeshan Yaseen ,
I am wondering what the manufacturer said. Since it all depends on the type of peptide. In general, this gives a pretty good guideline: “If the overall charge of the peptide is a positive value, you have a basic peptide. Initially try to dissolve the peptide in water. If the peptide does not dissolve, try 10% and higher solutions of acetic acid. If the peptide still does not dissolve, add TFA (
Thank you, @Rob Keller!
I was wondering that too why company hasn't sent guidelines with that package. Well, thanks for your answer. It does make sense, but making solution acidic for dissolving peptide won't affect the post-testing like antibiofilm and antimicrobial assay?
Please guide me about this too. Thanks.
Best,
Zeeshan
Dear Zeeshan Yaseen It depends on the 'magic' solution which will solubilise your peptide but if your peptide turns out to be hydrophobic then treatment with TFA (add it and then evaporate it with N2 flow) and later on solubilise with distilled water or an organic solvent like TFE (DMSO often works as well) might work. In such cases it should not be that much of a problem:
Your 1 mL end solution (I assume it will be some kind of buffered solution) only contains 64 uL of the peptide solution and 934 ul buffer.
How to treat the peptide solution, see for example:
Article Tryptophan fluorescence study on the interaction of the sign...
Best regards.
PS. You can minimise the potential influence of the peptide solution by making an even more concentrated peptide solution. 2 mg/mL would mean you add 32 uL (which is still doable).
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