Screen | NLRC4 Inflammasome Screen |
---|---|
Common Name | |
Posted On | 03/17/2017 3:20 PM |
Author | Hexin Shi |
Science Writer | Anne Murray |
Reagents and Solutions | |
Brewer’s thioglycolate medium, 4% 4% (w/v) Brewer’s thioglycolate medium powder (BBL Microbiology Systems, Cockeysville, MD) is added to distilled water pre-warmed to 37°C. Solution is autoclaved to sterilize and stored away from light. PEC recovery solution Hepes-buffered saline solution (Gibco, Invitrogen, Carlsbad, CA ) 5% (v/v) heat-inactivated fetal bovine serum (Atlanta Biologicals, Lawrenceville, GA) 200 IU/mL penicillin (Gibco) 200 mg/mL streptomycin (Gibco) PEC medium Dulbecco’s modified eagle medium (Mediatech Inc., Herndon, VA) 5% (v/v) heat-inactivated fetal bovine serum 200 IU/mL penicillin 200 mg/mL streptomycin MTT solution 5mg MTT (Sigma)/mL sterile PBS DMSO LPS (ALX-581-007-L002) Flagellin (adipogen, AG-40B-0095-C100) Lipofectamine 2000 (Invitrogen) Mouse IL-1b ELISA Ready-SET-Go (eBioscience: 88-7013-76) Wash Buffer 1X PBS, 0.05% Tween-20 Stop Solution 1M H3PO4 or 2N H2SO4 Pipettes 96-well plate (Corning Costar 9018 or NUNC Maxisorp (#44-2404)) 96-well ELISA plate reader DI water |
|
Method | |
Peritoneal exudate cell (PEC) isolation
Activation with LPS and flagellin
IL-1β ELISA
|