How to dissolve Neuro peptide galanin?
Answer:
This will sort of depend on the experiment in which the galanins will be used, for example, exposure of cells in culture or binding to isolated receptors. In either case, a working stock solution of 1 microgram per milliliter is useful, which is about 500 nanomoles per liter, so you can dilute it down to the nanomolar range by dilutions on the order of 10 microliters per milliliter, since the Kd's for most neuropeptides is on the order of 1 nM.
Now, for the buffer stock solution diluent. A safe bet is 50 mM HEPES, 1 mM magnesium chloride, 2.5 mM calcium chloride, and 1 mg/ml bovine or human serum albumin. The albumin will work as a carrier for the galanin and prevent adsorption to tube wall, etc. The divalent cations will enable a physiological conformation of the neuropeptide. The pH should be from 7.0 to 7.5, and 7.3 is a safe bet. Titrate the buffer before you add the peptide. If there is a major concern with oxidation of the cysteine sulfhydryl group, then add a very tiny amount of beta-mercaptoethanol, about 0.1 micromolar, or dithiothreitol (DTT), about 1 micromolar, to the buffer. Cell culture medium is already buffered with carbonate and phosphate so diluting your stock will not change the pH much. The HEPES buffer is a standard receptor affinity assay buffer.
The answers post by the user, for information only, FunQA.com does not guarantee the right.
More Questions and Answers: