3 replies to this topic

[thebigpiro]

My least favorite thing when running a western blot is trying to figure out what antibodies will work best. Loads of companies make loads of competing antibodies and it's very difficult to figure out which one is going to work the best without trying them. Well this is a pretty big community and we ought to be able to share good results with each other that will help save us time. So, here's a thread where (I hope) people will be able to quickly get feedback about which antibodies work best for their protein of interest. I'd like to see this thread take off and get pinned, but I guess we'll see if anyone else thinks this is a good idea. And, ah, if there's another thread like this already someone could point the way there for me....

I was recently testing a couple of GFP antibodies for mouse cardiac tissue. We have long used Abcam 290 but it was giving me a nonspecific band directly above the GFP band. So we tried another, Vector Labs BA-0702, which worked wonderfully well. I've attached a picture so you can see the difference. The picture isn't perfect but you can clearly see that the Abcam (top) has a nonspecific band, whereas the Vector Labs version (bottom) is clean.



So... if anyone would like to share decent iNOS or tubulin antibodies for use on mouse protein, I'd be much obliged. It sounds like tubulin is the way to go for a housekeeping protein if you're looking at ischemic tissue, so I need to check one out.

[laurequillo]

TUB 2.1, mouse monoclonal from SIGMA works quite well
GFP from Roche. It is a mouse one as well. The mixture of two clones, 7.1 and 13.1.

[Dwolfe38]

Personally, I think this is a GREAT idea for a thread.  I feel your pain, as lately it seems like 50% of the antibodies I obtain result in either no band, or are so non-specific as to be absolutely useless.  I find that there are certain companies that make a better product that others, with some being the absolute last resort.  It's unfortunate because a company like Santa Cruz makes Ab's to almost everything, but I've never gotten one that is clean (scratch that, their GAPDH is servicable.)  Anyway, my work involves a lot of Autophagic monitoring, to which I find that the LC3 from Novus is a good one, although over repeated uses, it seems to bind more to LC3-II than LC3-I.  Also for P62, both the Abnova antibody, and one by Progen is good as well.  I'm also a big fan of the Actin antibody from GenScript, although most Actin antibodies are decent (1:5000 of this blew out the signal).  

That said, I was hoping someone might be able to suggest antibodies for any of the following.

beta1 or beta2 adrenergic receptor proteins
CLC-7
Mucolipin 1

Thanks to anyone who can help.
-D

[Tom M]

has this thread died? Can it be resurrected?  
I'd like to know what antibodies work for people as well.  
GPCR antibodies especially.  Angiotensin antibodies specifically.  Anyone have any suggestions for these?
thanks.