HCC-1806
Origin:
Schulze Lab
Cell Medium:
DMEM/F12+10%FCS+Pen/Strep
Cell Seeding Density Per Well:
3000
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