Assay Overview

Plate Format: 384 well

Assay Cell Culture

RCC4

Origin: Swanton lab
Cell Medium: DMEM+10%FCS+1%PenStrep
Cell Seeding Density Per Well: 1250

Delivery Protocol - Reverse/Wet transfection

Protocol: Mix siRNA in 2.5uL HBSS to 375nM concentration. Mix 0.075uL Interferin in 3uL OPTIMEM. Mix together on plate and leave for 20 minutes. Add cells in 40ul/well. Top-up 50ul fresh media with drug per well after 24hour. Culture the cells for further 72hours
Delivery Reagent: Dharmafect3
Delivery Reagent Cat No: T-2003-07A
Delivery Reagent Supplier: Dharmacon
Delivery Reagent Concentration: 0.075ul/well
Silencing Reagent Concentration: 18.75nM
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