Identify Patient Zero of a Zombie Apocalypse With the Power of an ELISA!
Bio-Rad Curriculum and Training Specialist Tamica Stubbs tamica_stubbs@bio-rad.com Damon Tighe damon_tighe@bio-rad.com Sherri Andrews, Ph.D. sherri_andrews@bio-rad.com Leigh Brown leigh_brown@bio-rad.com
TV or real life? The Walking Dead show Feb 11,2014 Atlanta
Zombiefication Method of infection Symptoms Treatment Bodily fluid contact Airborne Foodborne Symptoms Undead Hungry for flesh Treatment None known
Introduction to Immunology, Antibodies and ELISA Fluid swapping and disease spreading! Indirect ELISA to narrow down who were the initial carriers of the disease
Crash Course in Immunology and ELISA Immune Response C. Macrophage A. Pathogen Zombie virus D. Macrophage B. B cells F. T cell E. Macrophage G. B cell J. Antibodies attach to pathogen Antibodies are ineffective Against Zombie Virus H. Memory B cells I. Plasma cells
Crash Course in Immunology and ELISA Antibody Structure ELISA tests are based on immune system antibody molecules Light chain Heavy chain Disulfide bonds Antigens
ELISA Enzyme-Linked Immunosorbant Assay (indirect) Add the purified antigen to all the wells. Incubate for 5 min. Rinse Add serum antibodies (student samples) to the appropriate wells. Incubate for 5 min. Rinse Add the enzyme-linked antibody to all wells. Incubate for 5 min. Rinse Add enzyme substrate to all wells. Incubate for 5 min.
Swapping Fluids and Disease Swap with 1 person at your table (record who) Add your 750 µl together, mix and each take 750 µl back
Swapping Fluids and Disease Swap with 1 person at the closest table (record who) Add your 750 µl together, mix and each take 750 µl back
Swapping Fluids and Disease Swap with 1 person at the another table (record who) Add your 750 µl together, mix and each take 750 µl back
Testing for who is infected ELISA Time! Testing for who is infected
ELISA Time! Testing for who is infected Your ID # Label Tubes
ELISA Time! Testing for who is infected Your ID # Add 50 µl “+” (Antigen)
Add 50ul “-” (wash buffer) Your ID # Your ID # Your ID #
Add 50ul Bodily Fluid Your ID # Your ID # Your ID #
ELISA Time! Testing for who is infected Your ID # Incubate: 5 minutes
ELISA Time! Testing for who is infected Your ID # Concept Corner
ELISA Time! Testing for who is infected Your ID # Concept Corner
ELISA Time! Testing for who is infected Polystyrene (plate) Protein (Antigen) Concept Corner Hydrophobic interactions
ELISA Time! Testing for who is infected Tap out liquid Your ID #
ELISA Time! Testing for who is infected Your ID # Wash with Buffer (contains Tween20)
ELISA Time! Testing for who is infected Tap out liquid Your ID #
ELISA Time! Testing for who is infected Your ID # Wash with Buffer (contains Tween20)
ELISA Time! Testing for who is infected Tap out liquid Your ID #
ELISA Time! Testing for who is infected Add 50 µl “PA” (Primary Antibody) Your ID # Your ID #
ELISA Time! Testing for who is infected Incubate: 5 minutes Your ID # Your ID #
ELISA Time! Testing for who is infected Your ID # Concept Corner
ELISA Time! Testing for who is infected Concept Corner Your ID # Your ID #
ELISA Time! Testing for who is infected Variable region Light chain Heavy chain constant region 3,368,600 antibody combos Concept Corner
ELISA Time! Testing for who is infected Tap out liquid Your ID #
ELISA Time! Testing for who is infected Wash with Buffer Your ID # Your ID #
ELISA Time! Testing for who is infected Tap out liquid Your ID #
ELISA Time! Testing for who is infected Wash with Buffer Your ID # Your ID #
ELISA Time! Testing for who is infected Tap out liquid Your ID #
ELISA Time! Testing for who is infected Add 50ul “SA” (Secondary Antibody) Your ID # Your ID #
ELISA Time! Testing for who is infected Your ID # Incubate: 5 minutes
ELISA Time! Testing for who is infected Your ID # Concept Corner
ELISA Time! Testing for who is infected Your ID # Concept Corner
ELISA Time! Testing for who is infected Your ID # Amplification of signal Introduce HRP (Horseradish Peroxidase) for colormetric detection Concept Corner
ELISA Time! Testing for who is infected Concept Corner Antigen = chicken gamma globulin Primary Antibody = anti-chicken polyclonal antibody Secondary Antibody = goat anti-rabbit conjugate to HRP
ELISA Time! Testing for who is infected Tap out liquid Your ID #
ELISA Time! Testing for who is infected Wash with Buffer Your ID # Your ID #
ELISA Time! Testing for who is infected Tap out liquid Your ID #
ELISA Time! Testing for who is infected Wash with Buffer Your ID # Your ID #
ELISA Time! Testing for who is infected Tap out liquid Your ID #
ELISA Time! Testing for who is infected Wash with Buffer Your ID # Your ID #
ELISA Time! Testing for who is infected Tap out liquid Your ID #
ELISA Time! Testing for who is infected Add 50 µl “Sub” (HRP substrate) Your ID # Your ID #
ELISA Time! Testing for who is infected Incubate: 5 minutes Watch for color change! Your ID # Your ID #
ELISA Time! Testing for who is infected Your ID # Concept Corner
ELISA Time! Testing for who is infected Concept Corner Your ID #
ELISA Time! Testing for who is infected Concept Corner
Deducing who was originally infected
Deducing who was originally infected Everyone Stand UP! If your ELISA was negative sit down
Deducing who was originally infected Round 1 If someone you shared with just sat down, also sit down Round 2 Round 3
Deducing who was originally infected Who is left?
Bio-Rad’s Immuno Explorer Lab What did we use today? Bio-Rad’s Immuno Explorer Lab 166-2400EDU
Other Zombie themed learning -> Texas instruments has a program for modeling (via graphing) the spread of a Zombie Virus: http://education.ti.com/en/us/stem-hollywood/zombies Bio-Rad’s Secrets of the Rainforest Kit can be used in a Zombie apocalypse story where students use column chromatography to isolate the Zombie Green Fluorescent Protein. https://www.dropbox.com/sh/zbjkffhfsmmpurw/AAAifY6D85YqRwWoBEGe__LGa?dl=0