Assay Overview

Plate Format: 384 well

Assay Cell Culture

PC-9

Origin: Downward Lab
Cell Medium: RPMI+10%FCS+1%PenStrep
Cell Seeding Density Per Well: 500

Delivery Protocol - Reverse/Wet transfection

Protocol: Mix siRNA in 2.5uL HBSS to 375nM concentration. Mix 0.075uL Dharmafect2 in 3uL OPTIMEM. Mix together on plate and leave for 20 minutes. Add cells in 80ul/well. After 48hrs, add drug by change media with 30nM erlotinib, culture the cells for 48hours.
Delivery Reagent: Dharmafect2
Delivery Reagent Cat No: T-2002
Delivery Reagent Supplier: Dharmacon
Delivery Reagent Concentration: 0.075ul/well
Silencing Reagent Concentration: 11nM
Silencing Reagent Supplier: Dharmacon
Silencing Reagent Origin: HTS Lab

Assay Treatments

Cell Fixation

Fixed in 80% Ethanol overnight
Ethanol
80%
long-term at -20C

Assay Staining

DAPI
1ug/ml
1 hour

Assay Readouts