I'm aware that inhibitory synapses are GABA and glycine and excitatory are glutamate. Gephyrin looks like a reasonable option for inhibitory - i.e. stains glycine and some GABA. Wondering if PSD-95 is genuinely excitatory specific?
Definitely could do with a protocol for these kinds of antibodies in 10-12 micron retinal sections if anyone could point me towards good articles or other resources.
you could try vGlut for excitatory.
Article Presynaptic inhibition of spinal sensory feedback ensures sm...
Hi,
Here is a good starting point in terms of protocol:
http://www.abcam.com/ps/pdf/protocols/double%20immunofluorescence%20-simultaneous%20protocol.pdf
and here:
http://research.jax.org/faculty/mills/protocols/immunofluorescence_frozen.pdf
Yes, gephyrin is good inhibitory marker for postsynaptic sites. I use rabbit anti-gephyrin from Abcam (ab25784), in 25µm thickness, 1:300 works very well.
Hope this help
Tahani
Hi all
Thanks a lot for the replies that's very helpful.
Tahani - what is your protocol for fixation when using Gephyrin? I have heard Gephyrin requires very low fixation time for retinal flat mounts (15-20 minutes only). Is this the same for fixation of retinal sections? Or is it just a standard fixation time with PFA 4%?
Cheers
Eamonn
I use the standard fixation we perfused the animal with 4%PFA for about 10 minutes. Then incubate it overnight with 4%PFA . Followed by incubating the brains in 20% sucrose.
If you have more questions I will be happy to help.
best
Tahani
- Badges
- Science topic