I hate Western blots. I think they are the most annoying, frustrating, and obnoxious experiments to do in lab. I hate that the process takes at least two days and that it's so antibody-dependent.
First, I am fully willing to admit that I am not good at Western blotting. My films usually look a little messy, and I've taken several steps to clean up my methods to no avail. The process is so touchy and there are so many steps that, apparently, I have trouble performing properly. That's my problem though.
What I hate about Westerns is how subjective they are. One antibody uses a milk block; another uses BSA; the last one takes three quarts of finely grated basil and a dash of kosher salt. The first one needs to be incubated overnight at 4 degrees; the second needs three hours at room temperature; the third needs to be set on a wooden stool in the midday sun from exactly noon to 2:30 PM.
The "voodoo" science involved in making these antibodies work bothers me.
It also doesn't help that the department's developer was made prior to the Cold War era (and I'm probably not even exaggerating too much).
What alternative is there? None. I may hate the process, but there's no way I'm getting around doing Western blots for the rest of my laboratory career.
Hear, hear!!
ReplyDeleteif is there a thing that i wanna kill, that is western blot :(
DeleteI found this short post today as I'm in the middle of a western blot and feeling pretty angry about it. :/ And most of all I hate the uncertainty of it! I have to wait until tomorrow after probing with my secondary to see if it even works... Ugh, at least we use fluorescent ab as our secondary, it is wayyy better than having to deal with chemiluminescence and dark rooms!
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