We lately had an interesting target for a mass-spec screen we wanted to follow and found out the THE experts for this protein sit right next door.

We had a meeting, they told us everything they do, they have a knockout mouse (yes!) and a good antibody (double yes!) that we could use.

We went back to the lab and tested the antibody on our tissue of interest, no signal...

Lets test the antibody on kidney tissue, it has been published that it is expressed there. We see a signal! It is different than what was described, but it is a staining.

Lets check in the knockout mouse, just to make sure. We see the same signal as in the wildtype.

We test a few more tissues. Nope.

Their antibody that they rely on for every staining they are doing is unspecific, sticks to everything that has a calcium-dependent lipid binding site and they never thought about making sure it is specific in their knockout animals that they had for yearssssss. WTF?

We learned our lesson. Every antibody is tested and then confirmed with an in-situ to see colocalization before we trust it.

thundara congrats on seeing the thing you wanted to see :)