anti rnf123 antibody picoband  (Boster Bio)

Journal: bioRxiv

Article Title: Identification of age-associated proteins and functional alterations in human primary retinal pigment epithelium cells

doi: 10.1101/2021.10.17.464744

Figure Lengend Snippet: Ubiquitin-mediated proteolysis in aging process of hRPE. (A) Western blotting (WB) analysis of the hRPE cells with different ages. RNF123, RNF149 and the ubiquitinated proteins in hRPE were up-regulated with age. (B) WB analysis of the ARPE-19 cells, pretreated with lutein for 6 h before stimulation with gradient concentrations of H 2 O 2 for 30 min. RNF123, RNF149 and the ubiquitinated proteins were up-regulated in ARPE-19 cells with increasing concentrations of H 2 O 2 , and this phenomenon can be reversed by lutein. (C) Cell viability of hRPE cells with different ages after H 2 O 2 stimulation. hRPE cells of O donors were more vulnerable to H 2 O 2 stimulation. (D) WB analysis of the O and the Y hRPE cells confirms the H 2 O 2 stimulation is related with the up-regulation of ubiquitylation of proteins. 67-year-old donor for the old, and 13-year-old donor for the young.

Article Snippet: 2.2 Compounds and Antibodies The antibodies and compounds used were commercially obtained: anti-RPE65 antibody (Abcam, #ab13826, 1:2000), anti-RNF123 antibody (BOSTER, #A09642-1, 1:500), anti-RNF149 (Bioss, #bs-9228R, 1:500), anti-ubiquitin antibody (Cell Signaling Technology, #3933, 1:2000), anti-β-Actin (Huabio, #EM21002, 1:5000), H2 O2 (Sigma-Aldrich, #323381), 40,6-diamidino-2-phenylindole (DAPI, Sigma-Aldrich, #9542), lutein (Sigma-Aldrich, #07168).

Techniques: Western Blot