Mouse anti sma cy3

Slides equilibrated to room temperature were subjected to heat-mediated antigen retrieval in 1× Target Retrieval Solution (Dako, S1699) by submerging the slides in the solution and steaming for 5 min and allowed to cool down. Sections were then washed three times in PBS with 0.1% Tween-20 (1× PBST) and incubated with primary antibodies diluted in DAKO Antibody Diluent (Cat No S0809, Agilent) at 4°C, overnight. Following washes in 1× PBST and incubation with secondary antibodies diluted in DAKO Antibody Diluent at room temperature for 1 h, slides were washed three times in 1× PBST and mounted with Mowiöl mounting medium. Primary antibodies used: rabbit anti-CHGA (1:500, Cat No 259 003, Synaptic Systems), rabbit anti-CYP11B1 (1:50, Cat No HPA056348, Sigma-Aldrich), mouse anti-SMA-Cy3 (1:1000, Cat No C6198, Sigma-Aldrich). DAPI was applied following staining at a concentration of 0.5 mg/ml. For primary antibody detection, secondary antibodies raised in donkey, conjugated with Alexa-488, and -555 fluorophores were used (1:1000, Molecular Probes, ThermoFisher Scientific).