AH08 Biochemical Assays for Bacterial Antigen Detection
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AH08 Biochemical Assays for Bacterial Antigen Detection Correct Answers and the Right Diagrams This study source was downloaded by 100000783208225 from CourseHero.com on 10-12-2022 10:11:48 GMT ... -05:00 https://www.coursehero.com/file/49944866/Lab-8-notebookdocx/ Objective: To understand the assays available for detecting bacterial or viral antigens (antibody generating pathogen). ELISA: Enzyme-Linked Immunosorbent Assay utilizes antibodies and colorimetric readout to indicate an antigen of interest. Western Blot: Separation of proteins based on size, utilizes antibodies and a colorimetric readout to indicate an antigen of interest. Procedure • Part 1: Loading the gel (SDS – dark purple color) • Part 2: Running the gel o 35 mAmps/gel • Part 3: Transferring the gel o 25 V for ~75 minutes • Part 4: Blocking the gel o 2% BSA for ~45 minutes for RT o Place on rocker • Part 5: Primary Antibody o TBST/BSA for ~45 minutes for RT o Place on rocker • Part 6: Secondary Antibody o TBST/BSA for ~45 minutes for RT o Place on rocker • Part 7: Membrane Development o BCIP/NBT at RT, covered o Place on rocker o Closely monitor Agglutination Test: A rapid assay to determine if antibodies to a specific pathogen are present. Most commonly test against patientserum blood. [Show More]
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