Could you suggest any good marker and corresponding antibody you employed to mark synapses? PSD95 seems a suitable option. I am using it on primary neuron culture from mouse cortex and hippocampus, but if it would work on cerebellar cultures would be also fine.
Thanks
francesco
SVP-38 (synaptophysin) is perfect marker for synaptic marker. Antibody SYP Antibody (SVP38): sc-12737 is good for bloting. For the presynaptic marker, Bassoon is good choice. Bassoon (D63B6) Rabbit mAb #6897 from cell signaling is OK for blots and IHC. For postsynaptic marker, as you know, PSD-95 is good marker. I remember 04-1066 Anti-PSD95 (NT) Antibody, clone EP2652Y is good for blots. I think these marker also work in cerebellar culture.
PSD95 from Neuromab (NeuroMab clone K28/43) is a good one. Meant to also mark PSD93 as well as -95 but despite this its a very reliable one and good for ICC and IHC. Synapsin I ( AB1543) also a good presynaptic marker alternative to synaptophysin.
I would recommend synapsin that would label the synaptic vesicle and therefore every mature presynaptic neuron.
I concur with Synaptophysin (>90-95% localisation on synaptic vesicles) as a good marker for the general presynaptic compartment and with Bassoon as a marker for the active zone.
Personally, I've had very good experience with the Synaptic Systems #101004 guinea pig antibody against Synaptophysin (http://sysy.com/products/s-physin1/facts-101004.php) and I find the Enzo SAP7F407 mouse monoclonal against Bassoon (http://www.enzolifesciences.com/ADI-VAM-PS003/bassoon-mab-sap7f407/) outstanding. Both antibodies are particularly good in ICC and IHC.
Good markers for the postsynapse include PSD-95 and Homer1.
The Synaptic Systems #160003 rabbit anti Homer1 (http://sysy.com/products/homer1/facts-160003.php) seems to be very good and colleagues of mine had good experience with PSD-95 antibodies from Cell Signaling.
For immunohistochemistry/immunofluorescence, I've had good luck with Synaptic Systems 106 001: a mouse monoclonal that recognizes 1a and 1b, and 106 002: a rabbit polyclonal specific for synapsins 1a/b and 2a/b. Both at 1:500
In addition to all of the previous suggestions, I've seen Shank3 used as a an alternative post-synaptic marker. Sometimes reviewers may ask for pre/post labeling with two independent antibodies, so having a few to choose from can be advantageous.
To stain GABAergic pre-synapses you can use VGAT. For glutamatergic pre-synapses I recommend vGlut2. Both work fine in vitro and in vivo. M
Thank you everybody, I think I am sticking for post-synaptic markers for the moment, as a colleague found some stored aliquoits of anti-synaptophysin.
The best way to visualize synapses is to use synaptophysin or synaptobrevin for presynaptic labeling and PSD-95 for postsynaptic one. The antibodies for the proteins are commercially available.
At Novus, we suggest the following markers for pre-synaptic, post-synaptic and synaptic vesicles analysis:
1. Synapsin I Antibody [NB300-104] pre-synaptic marker http://www.novusbio.com/Synapsin-I-Antibody_NB300-104.html
2. PSD-95 Antibody (6G6-1C9) [NB300-556] post-Synaptic Marker http://www.novusbio.com/PSD-95-Antibody-6G6-1C9_NB300-556.html
3. VAMP2 Antibody [NBP2-24599] Synaptic Vesicles Marker http://www.novusbio.com/VAMP-2-Antibody_NBP2-24599.html
This may be helpful :D
http://www.neuromics.com/ittrium/visit/A1x66x1y1x85b1x1x9cy1x6217x1x96y1x71bbx1y1x71b8x1x82
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