McClean: Alpha Factor Stock
This is the standard 1mg/ml stock solution we use for inducing the pheromone pathway in budding yeast. Generally, we find that the different batches of pheromone can be somewhat variable so it is a good idea to make up a stock that will last you through your experiment and keep it in your own -20°C box. If you have to switch stocks mid-experiment you need to do some calibration experiments to make sure the activity is the same.
- α1-Mating Factor acetate salt (Sigma T6901-1mg)
- DMSO (Fluka 41639, Ultra for molecular biology; stored in the Flammables cabinet)
Shake the new vial of α-factor to get all of the power on the bottom of the vial. Remove the lid and pipet 1ml of DMSO into the vial of α-factor from Sigma. Pipet up and down and put the lid back on an swirl to dissolve all of the alpha-factor into the DMSO. Then remove the liquid and aliquot it in 100μL eppendorfs into individual eppendorfs and store these in your own -20°C box.
Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input! *Megan N McClean: I generally find it useful to make a series 1:10 dilutions of the α-factor (1mg/m1, 100μg/mL, 10μg/mL, 1μg/mL) when I first make the stock, as these are the concentrations I generally start out with for most experiments. I usually make 900μL of all of these dilute stocks (ie, start with 100μL of 1mg/ml stock +900μL of DMSO, from that take 100 Please sign your name to your note by adding '''*~~~~''': to the beginning of your tip.
- Megan N McClean 14:01, 6 October 2011 (EDT)
or instead, discuss this protocol.