I tried glycine buffer but the yeast cells acidify the buffer so the pH goes down to 7-6 overnight. I need something that will stay around 9 and that isn't toxic to the cells.
To maintain a high pH (pH 9) for yeast cell suspensions, you can use a buffer that is effective in this pH range. Tris buffer is commonly used for maintaining alkaline pH and may be suitable for your needs. Here's how you can prepare Tris buffer at pH 9:
Tris Base (Tris(hydroxymethyl)aminomethane): Dissolve Tris base in distilled water to make a 1 M stock solution. The molecular weight of Tris base is 121.14 g/mol, so to make a 1 M solution, dissolve 121.14 g of Tris base in 1 liter of water.
Adjust pH: Adjust the pH of the Tris solution to 9 using concentrated hydrochloric acid (HCl) or concentrated sodium hydroxide (NaOH). Use a pH meter or pH strips to monitor and adjust the pH as necessary.
Final Dilution: Once the desired pH is achieved, dilute the Tris solution to the desired concentration for your experiment. Common working concentrations for Tris buffer range from 10 mM to 100 mM.
Sterilization: Filter-sterilize the Tris buffer using a 0.22 μm membrane filter to remove any particulate matter and microorganisms.
Storage: Store the Tris buffer at room temperature (if using within a few weeks) or at 4°C for longer-term storage. Avoid repeated freeze-thaw cycles.
Tris buffer is widely used in biological research and is generally compatible with yeast cell suspensions.