microtubule imaging - (Jul/07/2006 )

Hi All,

I'm doing some confocol microscopy, looking for a variety of proteins and small-molecule conjugates. Oddly, the only one that's not looking nice for me is tubulin. My primary has been around for awhile, so that could be the problem, or maybe I'm picking up depolymerized tubulin? I tend to just get the whole cell lit up.

Anyway, I was wondering if anyone had a favorite antibody for getting nice pictures of microtubules. There's so many to choose from! Is alpha better than beta?

Also, has anyone tried the various taxol-conjugates available from invitrogen? My cells are fixed and permablized, but these should pereferentially recognize the assembled structues, yes?

Other details: the cells are a variety of human cancer cells, including HeLa, T47D, and LNCaP.

Thanks in advance.

i'm a big fan of sigmas m anti alpha tubulin - always worked beautifully for me - in HeLa etc fixed in 4% PFA

Thanks, aussieuk

I'd looked at that one, but unfortunately I am stuck using a mouse antibody to another protein. I'm going to give the rat antibody from abcam a try - user reviews look good anyway.

By the way, I am currently using only 2% formaldehyde to fix my cells. Do you find 4% to be better?

Yes, the alpha tubulin from Sigma works very well no matter in western or tubulin imaging. I have tried.In fact,the alpha and the beta tubulin make up the microtubule.So the difference is the antibody from the different company.Good luck.

And of course, the -20 methnal is ok fixing the cell.

i'm pretty sure it is this one (but this was in my last lab, and i dont have the details with me now)
- it seems right from the specs on the sigma site
T5168