Screen Status: Primary
Screen Overview: Genome screen using ectodomain shedding assay
Screener: Marie Kveiborg (Parker Lab).
Extension:3505, Rm 203
Library: Full Genome 96
Library Description: 270 plates consisting of Kinome, GPCR, druggable and genome libraries
Library Supplier: Dharmacon
Library Format: 267 plate(s) in 96 well format
Replicate Info: 2 replicates
Technical replicates
Publications: The sorting protein PACS-2 promotes ErbB signalling by regulating recycling of the metalloproteinase ADAM17.. Dombernowsky et al., 2015
Keywords: ADAM proteases, HT1080, PKC
Cell Line: SJSA1

Screen Method

Treated

Day 1: Transfection of HT1080 cells stably expressing AP-HB-EGF

HT1080-AP-HB-EGF cells were reverse tranfected with siRNA using INTERFERin transfection reagent in 96-well plates.

Weeks 2 - 6: Immunofluorescent staining of cell surface expression of AP-HBEGF

Plates were stained with:

Weeks 2 - 6: Immunofluorescent staining of cell surface expression of AP-HBEGF

Plates were stained with:

Readout

Acumen Explorer eX3 laser scanning cytometer was used to determine cell number and cell surface expression of AP-HBEGF.