Parvalbumin Staining

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ilovebrains
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Parvalbumin Staining

Has anyone done immunocytochemistry for parvalbumin reactivity? I am trying to stain the GABA neurons of the medial septal region. The protocol I am using is only staining the cortex for some reason. However the samples I am using have been frozen in cryoprotectant and I am wondering if anyone knows anything about how quickly the parvalbumin protein degrades. Perhaps it degrades faster in the medial septum and fresh tissue would work better. I welcome any and all suggestions. Thank you.

sekirnjak
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ilovebrains wrote:Has anyone

ilovebrains wrote:

Has anyone done immunocytochemistry for parvalbumin reactivity? I am trying to stain the GABA neurons of the medial septal region. The protocol I am using is only staining the cortex for some reason. However the samples I am using have been frozen in cryoprotectant

I have successfully used a mouse antibody against parvalbumin (Sigma) at 1:100 on tissue that was fixed with 4% paraformaldehyde and 0.1% glutaraldehyde, no cryoprotectant used. How old was your tissue? An alternative would be to use anti-GAD antibodies to stain GABA-ergic neurons.