I have been given a protein of unknown concentration and have been asked to make a set of standards of a protein with a known concentration so we can quantify roughly via dot blot.The aim is to use a floursescent secondary antibody and to visualise via odessy.
I need to know how to work out the calculation part. I would like to learn both mass and molar.
For example, the information I have for my standard protein is 30mg/ml and 550uM. I want to put for example 1ul dots on the membrane for my blot. I might want several dilutions of my standard, for example ranging from 20pg to 100ng. How do I work it out to get varied concentrations from the stock to my 1ul volumes on my blot?
Also, should I do varied dilutions of the unknown protein?
Thanks, I'm so new to theis and very confused