Assay Overview

Plate Format: 96 well

Assay Cell Culture

HT-1080

Transgene: AP-HBEGF
Origin: Parker lab
Cell Medium: DMEM + 10% FBS + 1% L-Glutamine
Cell Seeding Density Per Well: 5000

Delivery Protocol - Reverse/Wet transfection

Protocol: Mix siRNA in 10uL HBSS to 37.5nM final concentration. Mix 0.3uL INTERFERin in 10uL OPTIMEM. Mix together on plate and leave for 20 minutes. Add cells in 80ul/well.
Delivery Reagent: INTERFERin
Delivery Reagent Cat No: 409-50
Delivery Reagent Supplier: Polyplus
Delivery Reagent Batch No: 03INF1011C
Delivery Reagent Concentration: 0.3ul/well
Silencing Reagent Concentration: 37.5nM
Silencing Reagent Supplier: Dharmacon
Silencing Reagent Origin: HTS lab

Assay Treatments

Cell Fixation

Fixed in 4% formaldehyde for 1 hour
Formaldehyde
4%
1 hour

Assay Staining

AP staining
5% BSA/PBS
DAPI
1ug/ml
1 hour

Assay Readouts