Can I dissolve freeze-dried catalase or lysozyme directly into de-ionised water or do I need to use a buffer (separate solutions, not both proteins in one)? I will not be storing the solutions and will be using them straight away.
If I need a buffer what should I use?
What would be the max concentration solution I can prepare?
Thank you
Catalase and lysozyme are enzymes, so the storage solution must maintain the protein's nature and activity
Thank you. I assume from your response that a pure water solution will not maintain the protein nature and activity. Is this true in all instances? Will working at low concentrations, for example, help to keep the protein intact? I am working on a new dehydration technique for proteins and would like to keep salts and additives out of the system as much as possible.
If not possible, what would you recommend as a dissolution buffer (sorry for the repetition but I have only worked with BSA to this point which I have dissolved in water)?
Pure water is fine if you are going to use it immediately. Lysozyme is a stable protein. There are about four disulfide linkages present in the native protein. So it does not unfold or denature very easily. And it should be fine for your purpose. But, if you want to be extra safe, the reccomended buffer is 10 mM Tris-HCl, pH 8.0. And reccomended maximum concentraton is 10 mg/ml.
However, I havent worked with catalases, so I am not sure about its solublity in water. But from this datasheet of a catalase kit, it is diluted in a buffer (10X buffer composition : 25 mM potassium phosphate, pH 7.5, containing 1 mM EDTA and 0.1% BSA)
Reference
https://www.caymanchem.com/pdfs/707002.pdf
Check the spec sheet: if the enzyme was lyophilised from a buffered solution, water will suffice. Otherwise, the manufacturer will suggest a suitable diluent.
In general it is preferable to keep the protein in buffer rather than water so that it keeps its stability, and to keep it more stable it is always recommended to be stored in a concentrated solution rather than diluted one, since the later one allow the protein to move freely which cause its denaturation and lose its activity & some time it is advisable to add low concentration of glycerol before storage in the freezer in order to reduce its movement and then keep it at a stable form.
The other thing is I would recommend using Phosphate buffer saline instead of Tris buffer since Tris buffer itself is unstable and you have to check its pH every day moreover it is subjected to bacteria growth, while PBS is much cheaper than the Tris furthermore it is stable >
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