We used to rely exclusively on the sc-52 rabbit anti c-Fos antibody from Santa Cruz to label free-floating coronal mouse and rat brain sections. My PI used it as a grad student, post-doc, and when he began his own lab and it worked great for DAB IHC and direct fluorescence. Unfortunately, we tried a couple of different LOTs recently and the results were terrible. To substitute, I have found that the Fast Fos protocol (search for this to find the paper that details the procedure) using sc-52G goat anti c-Fos works great. Good luck and let me know if you need any help.
We used to rely exclusively on the sc-52 rabbit anti c-Fos antibody from Santa Cruz to label free-floating coronal mouse and rat brain sections. My PI used it as a grad student, post-doc, and when he began his own lab and it worked great for DAB IHC and direct fluorescence. Unfortunately, we tried a couple of different LOTs recently and the results were terrible. To substitute, I have found that the Fast Fos protocol (search for this to find the paper that details the procedure) using sc-52G goat anti c-Fos works great. Good luck and let me know if you need any help.
As Justin and Francesco suggested, the sc-52 rabbit anti-Cfos from Santa Cruz Biotechnology works fine in many species. I used it at a concentration of 1:2000 on free-floating adult zebrafish brain sections. However, I also found that detecting cfos mRNA rather than Cfos protein is a more precise read-out of neuronal activity at least in fish.
I am sorry that I cannot make any suggestions for an anti-Arc antibody, but if you find a good one could you please be so kind to share your knowledge?
We were looking for an anti-c-fos outside of rabbit that would work in fish, and just had good success with Abcam's ab156802 in goat. The background is high, but the signal-to-noise ratio is at least as good as the lot of Santa Cruz rabbit anti-c-fos we have in the fridge. I did not find the current lot of SC goat anti-c-fos to work in our fish.
I am going to use the sc-52 from santa cruz. Does anyone have a good fixation method they use for brain tissue? I would like to avoid freezing. After overnight in 4% para, what should the tissue be stored in until slicing?
I'm also looking for a good c-fos antibody for perfusion-fixed mouse brains that is NOT the santa cruz antibody (I used to use in my grad school lab and it worked great, but my company won't let me buy from this vendor). If there are any suggestions of alternatives, it would be much appreciated. Thanks.
Although I used Santa Cruz regularly in the past with consistently good results for c-Fos, there are several reasons to not use them now, including the inconsistency and failure of lots recently.
I have recently used the Abcam ab102699 and it gave beautiful staining - although it stained so many more cells than I have seen with any other antibody, including a Millipore antibody I had stained the same tissue with previously, but they stopped making that particular one.
I have used the rabbit anti-cFos antibody from synaptic systems and it works like a charm on 4% PFA fixed rat brain free floating sections (no background). I used 1:100000 for DAB (sic!) and 1:20000 for fluorescent. I have also tried sc-52G and it works fine at 1:1000 for fluorescent, although a little more background.
We are getting a lot of nonspecific staining with santa cruz's sc-52 rbt anti-c-fos and are going to try alternatives. Debating whether to switch to the SC goat version, or try something non-SC. Justin, would you be willing to share your Fast Fos protocol?
I recommend to use a free antibody database available at labome.com. For antibodies to C-fos you can check the following link: https://www.labome.com/review/gene/human/c-Fos-antibody.html. In particular, Abcam has rabbit polyclonal c-Fos antibody (Abcam, ab7963) was used in immunohistochemistry. Santa Cruz Biotechnology offers mouse monoclonal (E-8) c-Fos antibody (Santa Cruz, sc-166940) used in immunohistochemistry. Cell Signaling Technology has c-Fos antibody (Cell Signaling Tech, 9F6) used in immunohistochemistry on mouse samples (Zhu et al., Nat Commun 2016). In addition, EMD Millipore has c-Fos antibody (Millipore, PC38-100UL) used in immunohistochemistry on mouse samples (ref: Hayashi et al, Science 2015).
For antibodies to Arc you can check the following link: https://www.labome.com/review/gene/human/ARC-antibody.html. Santa Cruz Biotechnology has mouse monoclonal (C-7) ARC antibody (Santa Cruz, sc-17839), which was used in immunohistochemistry - free floating section on mouse samples (ref: Gouty-Colomer et al., Mol Psychiatry. 2016).
I have used SC Santa Cruz Biotechnology monoclonal (E-8) c-Fos antibody (Santa Cruz, sc-166940) in 4% PFA perfused goldfish free floating sections. We have terrible results, both with DAB and fluorescence. We have tried also fast fos protocol.
Any suggestion about a cfos antibody (marking cells with a good signal/noise ratio) to use with teleost fish fixed free floating sections?
I am looking for an anti-cfos that is NOT raised in Rabbit (preferably not raised in mouse either) to use on rat free-floating 40 um brain sections. Do you know of any good-working antibodies?