How many times do I need to remake a stupid buffer before realizing that the buffer is too viscous not because I am incapable of making a 50% glycerol stock. No no, that would be too easy. It is to viscous because I am unable to calculate how many grams of NaCl I need to have 137 mM in 100ml.
That would O.8 grams not 8 grams, in case you were all wondering. Stupid freaking decimal points. That only wasted a day and half.
I then proceeded work on doing a particular IP the last time. I figured out all the issues with my western blot staining (my secondaries were no longer working). I was very diligent on the experiment.
- I calculated the protein concentration in my lysate
- I ensured that I took out 50 ug of protein to run in my control lane
- I calculated / double checked all my antibody concentration to ensure that it was all optimal for the pull down
- I ran my gel super slow to ensure nice clean bands
- I used fresh new antibodies solutions (took abs out of aliquots that I had shown work)
And if blogger was working properly right now, I would show you an lovely Licor western blog, nice sharp bands, beautiful specific binding. It would've have been a beautiful publication image.
Except for those damn bubbles in the transfer...grumble grumble grumble. Expletive, Expletive Expletive!
As a side note awesome PI thinks its hilarious and pointed out all the useful data we were able to get from the experiment. Which is positive. Then she told me how she had to redo a western 6 times to get it looking nice enough for publication....