Author: Gaikwad, Satish S.; Lee, Hyun-Jeong; Kim, Ji-Ye; Choi, Kang-Seuk
Title: Expression and serological application of recombinant epitope-repeat protein carrying an immunodominant epitope of Newcastle disease virus nucleoprotein Document date: 2019_1_31
ID: tr3ageky_13
Snippet: Proteins extracted from infected Sf9 cells were fractionated by 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis under reducing conditions. The separated proteins were blotted onto an Immun-Blot polyvinylidene fluoride membrane (Bio-Rad, Richmond, CA, USA) using a wet transfer system (Bio-Rad). The membrane was blocked with 5% skim milk in PBS containing 0.1% Tween 20 (PBST) at room temperature for 1 hour. After washing 3 times with .....
Document: Proteins extracted from infected Sf9 cells were fractionated by 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis under reducing conditions. The separated proteins were blotted onto an Immun-Blot polyvinylidene fluoride membrane (Bio-Rad, Richmond, CA, USA) using a wet transfer system (Bio-Rad). The membrane was blocked with 5% skim milk in PBS containing 0.1% Tween 20 (PBST) at room temperature for 1 hour. After washing 3 times with PBST, the blocked membrane was subsequently incubated with NDV chicken antiserum (diluted 1:100) for 1 hour, rinsed in PBST, and incubated with alkaline phosphatase-conjugated goat anti-chicken immunoglobulin (diluted 1:1,000, Pierce, Rockford, IL, USA) for 1 hour. Protein bands were visualized by enhanced chemiluminescence using NBT/BCIP solution (Sigma-Aldrich, St. Louis, MO, USA).
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