In lysis buffer, Sodium fluoride (NaF) act as a serine/threonine phosphatases inhibitor. I found different concentration as well as different storage condition depending on its application over cell and tissue as a important component of lysis buffer.
For your convenient answering, I am working on H9C2 cell lines. And, now I need to prepare sample by using lysis buffer to perform western blot.
So, I want to know that, which concentration and storage condition should I follow and why?
I would consider using 5-10 mM concentration, which is sufficient to inhibit protein phosphatases. Lysis buffer as well as cell lysate should be stored at -20 degree C.
I prepared NaF and stored at -20° C in small aliquote. Used at 1mM in fresh lysis buffer.
I suggested you to read material and methods of H9C2 works so you known the minimun concentration used with your cells and you can easily decide for the best.
In addition of what have been already said, keep the container use for storage tightly closed. Keep container in a cool, well-ventilated area.
Do not ingest. Do not breathe dust. In case of insufficient ventilation, wear suitable respiratory equipment. If ingested, seek medical advice immediately and show the container or the label. Avoid contact with skin and eyes. Keep away from incompatibles such as oxidizing agents, metals, acids, alkalis.
We used 1-5 mM as working concentration for various cell-lines like MCF7, MDAMB231, SKBR3 etc.
Along with these, we are using sodium ortho vanadate and PMSF as phosphatase inhibitors.
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