I am planning to do some WB from Drosophila heads. Because the major component of heads are the eyes, I would like to do WB only from brains by dissecting them. Does anyone have an idea how many you need for simple biological replica and the buffer that need to be used?
Thank you in advance
Luis De Navas Fernández
You can dissect the brains and put directly in 25 ul of cracking buffer (Sample Buffer Laemmly, 2X concentrate; Sigma S-3401 is the one I've used for larval imaginal discs). With 30 brains you should have enough for WB, but depending of the expression levels of the protein you are interested in maybe you will need to adjust a little bit this estimation.
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