ON-TARGETplus Human EPCAM (4072) siRNA - SMARTpool

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	siRNA concentration-30nM Replace medium 1h before transfection. Add diluted siRNA to siLentFect Reagent to make a mixture and incubate the mixture for 20 min at room temperature. Replace culture medium after 4h of transfection and incubate the plate at 37°C for 24 to 72 h.

Protocol tips
siRNA concentration-30nM Replace medium 1h before transfection. Add diluted siRNA to siLentFect Reagent to make a mixture and incubate the mixture for 20 min at room temperature. Replace culture medium after 4h of transfection and incubate the plate at 37°C for 24 to 72 h.

Publication protocol

DU145 human prostate cancer cells were obtained from ATCC (HTB-81) and maintained in high glucose DMEM with 10% fetal bovine serum (FBS), 1% glutamine, penicillin and streptomycin (GPS), and 1% sodium pyruvate (Invitrogen, Carlsbad, CA). Phase contrast microscopy was performed using a TE2000 microscope (Nikon) and RT SPOT camera with SPOT Advanced v4.0.9. software (Diagnostic Instruments, Inc., Sterling Heights, MI). Cells were transfected with siRNA using SilentFect (Biorad) in Opti-MEM I Reduced Serum Medium (Invitrogen), incubated for 4 hours, media changed, and cells used for assays at 48-72 hr. siRNAs were obtained from Thermo Scientific: ON-TARGETplus non-targeting control siRNA pool (D-001818-10-05), ON-TARGETplus human EPCAM siRNA pool (L-004568-01-0005), ON-TARGETplus human PLAU siRNA (L-006000-00-0005), ON-TARGETplus human ITGB4 siRNA pool (L-008011-00-0005). EPCAM and ITGB4 siRNAs were used at 30nM and PLAU siRNA used at 90nM for effective knockdown without toxicity.

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing siRNA / miRNA gene silencing Human - DU145 EPCAM using ON-TARGETplus Human EPCAM (4072) siRNA - SMARTpool from Dharmacon.

Manufacturer protocol

Download the product protocol from Dharmacon for ON-TARGETplus Human EPCAM (4072) siRNA - SMARTpool below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing siRNA / miRNA gene silencing Human - DU145 EPCAM using ON-TARGETplus Human EPCAM (4072) siRNA - SMARTpool from Dharmacon. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.