For a specific experiment, i am trying to avoid solubilizing calf skin collagen in acetic acid to have a solution around 5 mg/m. would 0.1 or 0.01 N Hcl work? does anyone has the experience and suggest what is the maximum solubility that is possible?
As I recall you will have to use 0.1 N HCl as well as Trypsin. See the dissolution procedure for crosslinked gelatin capsules (which are derived from pig/calf skin).
Without knowing the downstream assay purpose and conditions it is not possible and accurate to recommend solubility agents you may use in your buffer. There are many options which may increase the collagen solubility. For instance, do you need to follow an intact or digested (peptide mixture) form of collagen? Many proteases other than pepsin are inactive in acidic conditions. Chaotrpoes, salts, in some cases denaturants, or non-denaturing detergents can be preferred for protein solubilization. Other than acetic acid, it is possible to use HCl if the major concern is only lowering the pH. But I do not have any particular experience to increase collagen solubility by only changing the organic or inorganic acid type...
- Badges
- Science topic
- Similar topics
- Agriculture
- Horticulture
- Seeds