Author: Lee, Yu-Ching; Tsai, Keng-Chang; Leu, Sy-Jye; Wang, Tuan-Jen; Liu, Chia-Yu; Yang, Yi-Yuan
Title: Isolation, Characterization, and Molecular Modeling of a Rheumatoid Factor from a Hepatitis C Virus Infected Patient with Sjögren's Syndrome Document date: 2013_12_30
ID: 0zsn4lu3_10
Snippet: Briefly, microtiter plates were coated with 0.5 ug/well with human Fc fragment protein at 4 ∘ C overnight. The wells were blocked with 5% skim milk for 1 hr at room temperature. The expressed Fab molecules prepared as described above were distributed to wells in duplicate and incubated for 1 hr at room temperature. After washing with PBS containing 0.05% Tween-20 (PBST) six times, the wells were reacted with The Scientific World Journal 3 horse.....
Document: Briefly, microtiter plates were coated with 0.5 ug/well with human Fc fragment protein at 4 ∘ C overnight. The wells were blocked with 5% skim milk for 1 hr at room temperature. The expressed Fab molecules prepared as described above were distributed to wells in duplicate and incubated for 1 hr at room temperature. After washing with PBS containing 0.05% Tween-20 (PBST) six times, the wells were reacted with The Scientific World Journal 3 horseradish-peroxidase-(HRP-) conjugated goat anti-human or light chain antibodies (Jackson ImmunoResearch Laboratories) for 1 hr at room temperature. The wells were then washed with PBST six times and 3,5,5-Tetramethubezidine dihydrochloride (TMB) substrate was added for color development. Reaction was stopped by addition of 1 N HCl and absorbance was taken at 450 nm.
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