Journal: Investigative Ophthalmology & Visual Science
Article Title: Analysis of MTHFR, CBS, Glutathione, Taurine, and Hydrogen Sulfide Levels in Retinas of Hyperhomocysteinemic Mice
Figure Lengend Snippet: Analysis of retinal taurine levels and taurine transporter (TAUT). Retinas were harvested from 12- and 22-wk WT, Mthfr +/− , and Cbs +/− mice and were used to detect taurine levels using an amino acid analyzer and levels of TAUT by immunoblotting. ( A ) Quantitative data of retinal taurine levels analyzed as described in the text. ( B ) Representative immunoblot showing detection of TAUT. GAPDH was used as the internal control. There were no statistically significant differences between the groups from triplicate experiments.
Article Snippet: Membranes were blocked in 5% milk in TBST (1 hour at room temperature), followed by incubation with primary antibodies: goat anti-MTHFR (Abcam, Cambridge, MA, USA; 1:500), rabbit anti-CBS (Cell Signaling, Danvers, MA, USA; 1:250), rabbit anti-CTH (Thermo-Fisher Scientific, Waltham, MA, USA; 1:500), rabbit anti-3MST (Aviva Systems Biology, San Diego, CA, USA; 1:500), rabbit anti-xCT (1:800), and rabbit anti–taurine transporter (TAUT) (Alomone Labs, Jerusalem, Israel; 1:200) in 5% milk in TBST buffer at 4°C overnight.
Techniques: Western Blot