Lab Lesson of the Day: Check Your pHs

All last week I was trying to optimize my Western Blot procedure. This is what Westerns are supposed to look like:


This is what my Westerns looked like:



I was doing all of these elaborate set ups to figure out the problem. I tried other labs antibodies, I tried other labs blotting apparatuses, I tried other labs membranes, I tried different blocking agents, nothing was working! I mean, this is a really simple procedure, and I was just trying to get a cellular control (actin) to work! There was no reason for this to be so hard! I could teach a 10 year old to do this protocol (well, not 'me' technically because mine were screwing up, but still).

I figured out what the problem was today. I was mixing some other solutions, and I realized I didnt check the pH of my PBS. Its supposed to be 7.4. I measured mine: 4.7.

4.7.

*blink*

AAAAAAAAAARRRRRRRRRGGGGGGGGGG!!!

If you dont work with chemicals much and dont know what that means, think of it this way-- Its like trying to grow an oak tree on Mars. It just aint gonna work right.

So I tell my boss what the deal was. He was speaking with a couple other professors at the time. He turned to them and said, "ERV designed some wonderful experiments last week to diagnose why our Western Blots werent working, and she figured out what the problem was today. (turning to me) But thats not going to happen again, right?"

Hes a good boss :)