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Our Abpromise guarantee covers the use of ab18736 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use on perfusion fixed tissue (1% sodium Nitrite in 0.1M TBS, pH, 7.4 for 20 min then 4% formaldehyde containing 15% picric acid in 0.1M TBS,pH, 7.4 for 20min). Please note sections must be post fixed if the tissue is cut freshly.
|IHC-P||Use at an assay dependent concentration.|
|ICC/IF||1/100. PubMed: 18393628|
ab18736 staining Choline Acetyltransferase in mouse brain tissue sections by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue samples were fixed by perfusion with paraformaldehyde, permeablized with 0.3% Triton X-100 and blocked with 3% donkey serum in 0.3% Triton X-100/TBS for 1 hour at 21°C. The sample was incubated with primary antibody (1/1000 in 1% donkey serum + 0.3% Triton X-100/TBS) at 4°C for 24 hours. An Alexa Fluor® 594-conjugated donkey anti-sheep polyclonal (1/200) was used as the secondary antibody.
Immunofluorescence of Rat Forebrain tissue labeling ChAT with ab18736 using a 1µg dilution.
Immunohistochemical detection of ChAT in rat basal forebrain using Sheep anti ChAT (ab18736).
Rat was perfused with 1% sodium Nitrite in 0.1M TBS, pH, 7.4 for 20 min. and then fixed with 4% formaldehyde containing 15% picric acid in 0.1M TBS, pH, 7.4 for 30 min. The brain was then cryoprotected in 30% overnight and cut into 40 nm free-floating sections. Sections were washed 2x with TBS for 20 min. and passed through asending ethanol and finally transferred to 0.1 M TBS for rehydration for 1h.
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