Related topics: Alphabetical list, Developmental biology, Neurotransmission
On April 28, 2020
Justin Botterill, The Nathan S. Kline Institute for Psychiatric Research wrote:
Immunostaining were done on 4% paraformaldehyde perfusion-fixed brains that were cut at 50µm on a vibratome.
Antibody specificity appears fine and is consistent with literature. We used 1:1000 concentration. Some cells are incredibly well stained and others are only moderately stained, requiring you to make a inclusion criteria for counts. Background signal can be high -which can be challenging to optimize reaction.
Signal is improved following antigen retrieval (ph 6.0 sodium citrate buffer @ 85-95 celsisus for 30 minutes).
For more information see Botterill et al. Cell Reports 2019, “An Excitatory and Epileptogenic Effect of Dentate Gyrus Mossy Cells in a Mouse Model of Epilepsy”