TransIT-TKO Transfection Reagent

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Prepare TransIT-TKO Reagent:siRNA complexes in serum- free growth medium, preferably optiMEM.

Upstream tips
Prepare TransIT-TKO Reagent:siRNA complexes in serum- free growth medium, preferably optiMEM.

Publication protocol

Transfection with IL-1β/GFP or GFP expression plasmids
The plasmid pEGFP-C3 encoding wild-type GFP optimized for brighter fluorescence and higher expression in mammalian cells was obtained from Clontech Laboratories, Mountain View, CA. The plasmid RG202079 in pCMV6-AC-GFP vector designed to express IL-1β in mammalian cells as a GFP-tagged protein was obtained from Origene (Rockville, MD). HEK293 cells and human astrocytes were transfected with pEGFP-C3 or RG202079 (IL-1B - GFP) plasmids in 96-well plate or 6 cm dishes using TransIT-TKO transfection reagent (Mirus BioLLC, Madison, WI) or Continuum™ transfection reagent (Gemini Bio-Products, West Sacramento, CA) following the manufacturer’s instructions. After 24 hr, GFP was detected in 96-well plate using Olympus fluorescence microscope. The cells from 6 cm dishes (after control for GFP expression by fluorescence) were scrapped into PBS buffer, lysed, and used for Western blotting.

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Manufacturer protocol

Download the product protocol from Mirus for TransIT-TKO Transfection Reagent below.

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