Plate Format:

96 well

184B5

Origin:	Schulze Lab
Cell Medium:	DMEM/F12+10%FCS+Pen/Strep
Cell Seeding Density Per Well:	5000
Cell Culture Conditions:	0.5%O

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. Change the media with 100ul fresh media after 24hours, then culture the cells for further 72hours
Delivery Reagent:	Lullaby
Delivery Reagent Cat No:	LL73000
Delivery Reagent Supplier:	Oz Bioscience
Delivery Reagent Concentration:	0.1ul/well
Silencing Reagent Concentration:	37.5nM
Silencing Reagent Supplier:	Dharmacon
Silencing Reagent Origin:	Schulze Lab

Description:Fixed in 80% Ethanol overnight
Fixation Compounds:Ethanol
Compound Concentration:80%
Treatment Time:long-term at -20C

Description:Cleaved caspase-3 staining
Blocking:3%Donkey serum/PBS
Permeabilisation:1hour

• Primary Antibody - Cleaved caspase-3 (Asp175) antibody, Rabbit polyclonal
  Supplier:Cell Signalling (NEB)
  Cat No:9661
  Batch No:Lot18
  Dilution:1/750
  Incubation Time:2hour
  
• Secondary Antibody - Alexa Flour 488 donkey anti-rabbit IgG secondary antibody
  Supplier:Molecular Probes
  Cat No:A21206
  Batch No:913921
  Dilution:1/1500
  Incubation Time:1 hour
  

Nuclear Staining:DAPI
Nuclear Staining Concentration:1ug/ml
Nuclear Staining Incubation Time:1 hour

• TTP Acumen Explorer Ch1
  Localisation:nucleus
  Stain:DAPI
  
• TTP Acumen Explorer Ch2
  Localisation:nucleus
  Stain:Caspase-3