Anti phosphor stat3 tyr705

The samples of the periodontal tissue and cerebral cortex of rats in four groups were homogenized and lysed by RIPA containing 1% protease inhibitor cocktail and 1% PMSF (Beyotime, Shanghai, China). Equal amounts of protein were separated by SDS polyacrylamide gel electrophoresis and transferred onto PVDF membrane blocked with 5% skimmed milk as previously described [19 (link)]. A pre-stained protein marker (Thermo Fisher Scientific, MA, USA) was run in parallel to detect the molecular weight of proteins. GAPDH was used as a protein loading control according to its high and constant expression in most tissues and cell types, earning the gene and its protein housekeeping status. The proteins were probed with appropriate antibodies including anti-JAK2 (Rabbit mAb, 1:500, 3230 T; Cell Signaling Technology, USA), anti-STAT3 (Rabbit mAb, 1:1000, 4904 T; Cell Signaling Technology, USA), anti-phosphor STAT3-Tyr705 (Rabbit mAb, 1:200, 9145S; Cell Signaling Technology, USA), and anti-GAPDH (Rabbit mAb, 1:1000, AB-P-R001, Goodhere Biotechnology Co., Hangzhou, China). The data were quantified by ImageJ 1.51j8.