Assay Overview

Plate Format: 96 well

Assay Cell Culture

MCF-7

Origin: Schulze Lab
Cell Medium: DMEM+10%FCS+Pen/Strep
Cell Seeding Density Per Well: 2000
Cell Culture Conditions: 10%CO2

Delivery Protocol - Reverse/Wet transfection

Protocol: Mix siRNA in 10uL HBSS to 37.5nM final concentration. Mix 0.1uL Lullaby in 10uL OPTIMEM. Mix together on plate and leave for 20 minutes. Add cells in 80ul/well. Top-up the media with 100ul fresh media after 24hours, then culture the cells for further 72hours under normal/hypoxia condition
Delivery Reagent: Lullaby
Delivery Reagent Cat No: LL73000
Delivery Reagent Supplier: Oz Bioscience
Delivery Reagent Batch No: LL101-214
Delivery Reagent Concentration: 0.1ul/well
Silencing Reagent Concentration: 37.5nM
Silencing Reagent Supplier: Dharmacon (On-target non-target plus)
Silencing Reagent Origin: Schulze Lab

Cell Fixation

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

Assay Staining

DAPI
1ug/ml
1 hour

Assay Readouts