Journal: bioRxiv
Article Title: Development and characterization of phospho-ubiquitin antibodies to monitor PINK1-PRKN signaling in cells and tissue
doi: 10.1101/2024.01.15.575715
Figure Lengend Snippet: Assessing the top p-S65-Ub antibodies in ELISA using recombinant protein. ( A ) Sandwich ELISA results for the detection of recombinant K48 (solid line) and K63 (dash line) p-S65-Ub tetramers (Ub4) that were serially diluted to the range of 1–1,000,000 pg/ml. ( B ) Direct ELISA results for the detection of p-S65-PRKN and nonphosphorylated PRKN recombinant proteins that were serially diluted to the range of 1–1,000,000 pg/ml. N = 2. Data are shown as mean with SD.
Article Snippet: Recombinant p-S65-PRKN (Boston Biochem, E3–166), non-phosphorylated PRKN (Boston Biochem, E3–160) and p-S65-Ub monomer (Boston Biochem, U102) were initially dissolved in RIPA buffer (stock concentration of 1 μg/μl), aliquoted in 1 μl aliquots, flash-frozen in liquid nitrogen and stored at −80°C.
Techniques: Enzyme-linked Immunosorbent Assay, Recombinant, Sandwich ELISA, Direct ELISA