Assay Overview

Plate Format: 384 well

Assay Cell Culture

MCF-10A

Origin: Downward lab
Cell Medium: E4/F12+5%horse serum+20ng/mlEGF+100ng/mlCholera toxin+10ug/ml Insulin+0.5ug/mlHydrocortisone+PenStrep
Cell Seeding Density Per Well: 3000

Delivery Protocol - Reverse/Wet transfection

Protocol: Mix siRNA in 10uL HBSS to 37.5nM final concentration. Mix 0.225uL Lullaby in 10uL OPTIMEM. Mix together on plate and leave for 20 minutes. Add cells in 80ul/well. Top-up 100ul fresh media after 48hours, then culture the cells for further 48hours
Delivery Reagent: Lullaby
Delivery Reagent Cat No: LL73000
Delivery Reagent Supplier: Oz Bioscience
Delivery Reagent Batch No: LL101-214
Delivery Reagent Concentration: 0.225ul/well
Silencing Reagent Concentration: 37.5nM
Silencing Reagent Supplier: Dharmacon
Silencing Reagent Origin: HTS Lab

Cell Fixation

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

Assay Staining

DAPI
1ug/ml
1 hour

Assay Readouts