Can anybody recommend a good antibody for immunohistochemistry in perfused rat brain against VMAT2? we have been using for years the old Chemicon Ab 1767, now not sold any more. Anybody can recommend one that works nicely on their hands?
Marina,
See datasheets of your antibody and search the name of the clone (call the Chemical office if not written on datasheets).
After search on the web (biocompare or other antibody compare site) your antibody.
In France, Merck Millipore has the Chemicon antibody reference AB1598P.
See:
http://www.merckmillipore.com/DK/en/product/Anti-Vesicular-Monoamine-Transporter-2-Antibody,MM_NF-AB1598P
Good luck,
VM
Hopefully others will be able to offer you personal recommendations of good anti-VMAT2 Abs. We can not vouch for any particular Ab, but if you need, we can help you locate a number of possible new sources. Just search here http://www.linscottsdirectory.com/search/antibodies for VMAT2, and select Rat and IHC from the Reactivity and Applications filters.
Thanks for the helpful comments, I will check them all. However, I was more on the search of any personal recommendation of and antibody that could have already worked for any other researchers. So please, If so just let me know.
Dear all, I found one good antibody: Goat anti-VMAT2 Everest Biotech SLC18A2
Excellent in rat free floating sections from PFA perfused brains at 1:5000. Enjoy!
Hi Marina,
I just used the Everest antibody (recently ordered batch) in mouse striatum cryosections (1:200) and in cultured dopaminergic neurons. At 1:200 there was a lot of unspecific staining in the sections. Perhaps higher dilution as you used will work, but in the cell cultures the staining looked unconvincing. I double labelled for TH and although there was clear axonal staining with the Everest antibody, it was in TH unlabeled axons. Marisela Morales' lab used this antibody for a very nice study in Nature Neuroscience recently. So I'm a bit puzzled, perhaps I got a bad batch. Previously, i used the one from Phoenix Pharmaceuticals, which worked beautifully for IHC (but see western blots in MM's paper!), but it is not available right now - they are still working on making another batch that works. Do you use any special fixation protocol (other than 4% paraformaldehyde)?
Yvonne, my suggestion is dilute it, this antibody has a good signal at 1:5000 and we are now testing even more diluted, so I believe 1:200 is extremely high.
I am not sure how was your preparation but TH is a protein highly regulated and can suddenly become undetectable in Cathecolaminergic cells . So, my suggestion is to be cautious with that.
In any case, it works fine for use. Please see:
http://pabmabs.com/wordpress/?p=4033
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