Western blotting is the bread and butter of many a molecular biologist. It is also frequently the most loathed of all techniques. Last week, it was working just fine. But this week? No such luck, but WHY?!
1. You did it on a different day than last time
Last week, you set everything up to run on Tuesday at 10AM. This week, it was Wednesday at 2PM. Western’s know what day of the week it is. They mark it off on their calendar. If it isn’t the same day, try again next week.
2. You had the radio on
Did you have the radio on last week? Did you change the station? Was it off during primary incubation, but on during secondary? The membrane KNOWS. Don’t underestimate the membranes love of music. This could be the difference between a band or no band on that blot.
3. You wore a different outfit
Westerns are notorious fashion critics. Pairing those shoes with that lab coat? Not as good as last weeks sartorial choices. Go back to the wardrobe and try again. Surely you have a set working wardrobe to ensure you only wear the same clothes on the same day each week? This isn’t amateur hour.
4. Didn’t sing to your cells?
Please tell me you didn’t make this mistake? People, puh-lease. Your cells favourite genre will vary will dependent on their species, and bodily origin. My breast cancer cells are partial to a bit of dance music. No Tiësto? Don’t expect to see increased turnover.
5. You stood in a different position
Remember last week? You stood to the left of your lab bench. This week, that side is full of things you haven’t tidied up. Don’t make the mistake of standing in a different position. Nothing will work. Did you run your gel on a different bench? You may as well stop there. It’s almost definitely not going to work.
6. You didn’t show enough enthusiasm
Get the cheerleaders out, tell the ECL that you believe in it. It’s kind of like fairies. If you don’t truly believe, you’ll never see it in real life. Have some faith, and maybe a little bit of pixie dust wouldn’t hurt, either.
7. The sky is cloudy
Last week’s glorious sunshine is a distant memory. Get used to this weeks dull, grey skies. Wait, you didn’t realise the outside weather would alter your (indoor, temperature controlled) experiments? Think again.