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I have just some trouble staining synapses using synaptophysin. I would like to combine synaptophysin and tyrosine hydroxylase like they're an active synapse on a target cell but it seems that synaptophysin and tyrosine hydroxylase won't colocalize. I'm working on fixed and cryocut rat brain sections.
Do you have any suggestions?
P.S. I am a master student and I don't have a lot of practical skills.
Thank you
S.
Mon Aug 22, 2011 2:50 pm
Carl
Rank: Member
Joined: Oct 11, 2004
Posts: 1871
Post subject: Hi Save...welcome.
What primary Abs are you using?
In the image gallery are many images of P.wax TH and synaptic markers which also work very well on frozen sections.
However, I would not expect a synaptic marker to co-localise with TH.
I may be wrong...
Do you obtain positive results with these 2 Abs ( not co-localised)?
Curious Carl
Mon Aug 22, 2011 5:11 pm
Save
Rank: Member
Joined: Aug 22, 2011
Posts: 2
Post subject:
Hi Carl.
I'm using an a-TH from bovine adrenal medulla in rabbit with the second antibody from Jackson laboratory, Donkey a-Rabbit Cy3. I think also that this aliquot is older than me..
Until now I have positive results on both of them. I'll go now to see the images you are talking about.
Thank you
S.
Tue Aug 23, 2011 7:22 am
jan
Rank: Member
Joined: Jul 07, 2008
Posts: 21
Post subject:
I have experience with both TH as well as synaptophysin. Carl is right, why would you expect double labelling? TH is in the cell body of the dopaminergic cells whereas SP is in the synapses. You might see some overlap but the bulk of reaction should be separate.
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