Antibody staining for dendritic/spine retraction

kristie

Hi all, I was just wondering if anyone had any suggestions for me for a protein to stain for looking at dendritic spine retraction in a stress protocol (or any other protocol that induces changes). I have some structural protein ideas in mind, but I am just checking to see if anyone could suggest a protein present/expressed when alterations dendritic morphology occurs (I'm a little new to this field).

Cheers.

TomPhillips

Hi, MAP2 is a good target for measuring dendrites and should work in Clarity

kristie

Thanks. I was thinking MAP2 might work

helen

I've been trying MAP2, and while i get staining, it's hard to resolve structure above the background. If you get it working i'd love to know your technique.

kristie

helen said:

I've been trying MAP2, and while i get staining, it's hard to resolve structure above the background. If you get it working i'd love to know your technique.

Hi Helen, I went for Phalloidin stain (F actin) to try and target dendritic spines and unfortunately I'm having a lot of issues with being able to distinguish our targets from the background. I increased the concentration of the stain and left it for longer, and had a longer wash time, but while I am getting more definition in the staining, I'm still having issues with the background.

Were you also targeting dendrites? If not what was the aim of your stain?
Have you had any luck?