Twenty-four hours after TAMRA-MPH treatment, cells were fixed by incubating for 15 min in 4% paraformaldehyde with 4% sucrose in 1 M PBS, pH 7.4. Slides were incubated for 1 h at room temperature in blocking solution (2% w/v bovine serum albumin (BSA, Sigma-Aldrich) plus 3% v/v normal goat serum (Sigma-Aldrich) in PBS, then overnight at 4°C with Synapsin III antibody (1:600, Synaptic System). On the following day, cells were incubated for 1 h at room temperature with the 488 fluorochrome-conjugated secondary antibody (Alexa Fluor® 488, Jackson Immunoresearch) diluted in 0.1% Triton X-100 PBS plus BSA 1 mg/ml. After three washes in 0.1% Triton X-100 PBS, cell nuclei were counterstained with Hoechst 33258 dye (Sigma-Aldrich), and the coverslips were mounted onto glass slides using Vectashield (Vector Laboratories, Burlingame, CA).
Synapsin 3 antibody
Manufactured by Synaptic Systems
The Synapsin III antibody is a research tool used to detect and analyze the presence of Synapsin III, a protein involved in the regulation of neurotransmitter release at synaptic terminals. It can be used in various techniques, such as Western blotting, immunohistochemistry, and immunocytochemistry, to study the expression and localization of Synapsin III in biological samples.
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Lab products found in correlation
2 protocols using synapsin 3 antibody
1
Immunofluorescence Staining of Synapsin III
2
SYN1/2/3 Triple Knockout Mouse Brain Profiling
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Cortices from wild type or SYN1/2/3 triple knockout mice (TKO) were homogenized in RIPA buffer, and lysates were used for SDS-PAGE and Western blotting as described (Höltje et al., 2017 (link)). Membranes were incubated with CBA-positive sera (= CBA score 2) diluted 1:200. For Western blotting of transfected HEK293 cells, transfected cells were grown on 6-well plates and homogenized in phosphate-buffered saline (PBS) with protease inhibitors. Homogenates were used for SDS-PAGE and Western blotting. Untransfected cells served as control. A rabbit polyclonal synapsin-I/II antibody (Synaptic Systems, #106002) served as positive control to detect synapsin isoforms in brain lysates and transfected HEK cells. A monoclonal glyceraldehyde-3-phosphate dehydroxygenase antibody (GAPDH, Merck Millipore, #MAB374) was used as loading control.
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