# Best antibodies to label different neuronal populations **URL:** \ **Category:** Microscopy (IHC/ISH/ICC/In Vivo)\ **Created:** [October 16, 2017, 3:04pm UTC](https://forum.painresearcher.net/t/best-antibodies-to-label-different-neuronal-populations/304 "2017-10-16T15:04:54Z")\ **Posts on this page:** 1\ **Showing post:** 2
**Author:** ![achamess](https://yyz2.discourse-cdn.com/flex030/user_avatar/forum.painresearcher.net/achamess/32/534_2.png) [@achamess](https://forum.painresearcher.net/u/achamess)\ **Post date:** [October 16, 2017, 3:56pm UTC](https://forum.painresearcher.net/t/best-antibodies-to-label-different-neuronal-populations/304/2 "2017-10-16T15:56:03Z")
The problem with antibodies for labeling neurons is that the best markers are not well expressed in neuronal somata. So for example, the best excitatory marker is Slc17a6 (VGLUT2). Antibodies to VGLUT2 mark terminals, however. Similarly, for inhibitory neurons, GAD1/2 or VGAT are best, but they too are not expressed in the soma. So, you have two choices. Either you do in situ hybridization. See this post for an example of VGAT (pan-inhibitory) > [@RNAscope (In situ hybridization) to study gene expression in pain-related tissues](http://forum.painresearcher.net/t/rnascope-in-situ-hybridization-to-study-gene-expression-in-pain-related-tissues/102): > > In situ hybridization is a powerful method to interrogate gene expression in tissues. It is known as being a challenging procedure due to the many steps where errors and obstacles arise. We’ve recently begun to use a commercial system called [RNAscope](https://acdbio.com/science/technology-overview) from ACD Bio that gets around many of the pitfalls of traditional ISH. I’ve used it a lot now in the spinal cord and DRG. Below is an image of the gene VGAT (Slc32a1) in the dorsal spinal cord. There is nice expression, and most importantly, it was… ISH is great but a pain in the butt to do traditionally, or expensive with RNAscope. Your other option is to use transcription factor antibodies. In particular: Pax2 = Inhibitory. This one works (1:500). > **[PAX2 Polyclonal Antibody (71-6000)](https://www.thermofisher.com/antibody/product/PAX2-Antibody-Polyclonal/71-6000)** > > Invitrogen Anti-PAX2 Polyclonal, Catalog # 71-6000. Tested in Western Blot (WB), Immunohistochemistry (IHC), Immunoprecipitation (IP) and ELISA (ELISA) applications. This antibody reacts with Human, Mouse, Ovine, Rat, Xenopus laevis, Zebrafish... For excitatory in the dorsal horn, TLX3 is used. Unfortunately, the widely used TLX3 antibody is custom from Carmen Birchmaeir’s lab in Germany. I don’t know about commericial. I made a panel like this here: > [@Stuff I tried during grad school/post-doc that won't get published but could be useful](http://forum.painresearcher.net/t/stuff-i-tried-during-grad-school-post-doc-that-wont-get-published-but-could-be-useful/302/6): > > I talked about this in another thread Here are some more images. Overall, this FosGFP line is good for activity-dependent labeling. It’s not perfect. There are some Fos+ (IHC) neurons that don’t get labeled, but overall, it’s useful. Could be good for electrophysiology. The stimulus here was 55ºC heat for 30s, followed by perfusion 2 hours after the stimulus. [fos-slices-panel-01-01] Here is a recent paper doing the same: > **[A Brainstem-Spinal Cord Inhibitory Circuit for Mechanical Pain Modulation by...](https://pubmed.ncbi.nlm.nih.gov/28162807/)** > > Pain thresholds are, in part, set as a function of emotional and internal states by descending modulation of nociceptive transmission in the spinal cord. Neurons of the rostral ventromedial medulla (RVM) are thought to critically contribute to this... Good luck! --- _[View the full topic](https://forum.painresearcher.net/t/best-antibodies-to-label-different-neuronal-populations/304)._