ON-TARGETplus Rat Fyn siRNA

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Seed 3×10^3 cells/well into 48 well tissue culture plates.	After 24h treat cells with 400 nM Fyn siRNA for 4h. Following incubation, add growth medium containing 3 times the normal concentration of serum without removing the transfection mixture. Incubate cells for 72 hours.

Upstream tips
Seed 3×10^3 cells/well into 48 well tissue culture plates.
Protocol tips
After 24h treat cells with 400 nM Fyn siRNA for 4h. Following incubation, add growth medium containing 3 times the normal concentration of serum without removing the transfection mixture. Incubate cells for 72 hours.

Publication protocol

To verify the role of Fyn in GDNF signaling, knockdown of Fyn was performed using Fyn specific ON-Target plus SMARTpool siRNA (Dharmacon, Thermo Scientific, Lafayette, CO). Targeted RNA sequences included UCGAACGCAUGAAUUAUAU, GUAGUUCCCUGUCACAAAG, AGAGGUACCUUUCUUAUCC, and UGACCUCCAUCCCGAACUA. Briefly, sensory and motor-derived SCs were seeded at 3×103 cells/well into 48 well tissue culture plates in ABAM-free media. After 24 hours, media was removed and SCs were treated with 400 nM Fyn siRNA with oligofectamine reagent (Invitrogen) in Opti-MEM and ABAM-free, serum-free DMEM media for 4 hours. Control SCs were not treated with Fyn siRNA. After 4 hours, cells were given DMEM media supplemented with 3x normal FBS alone or with 100 ng/mL GDNF. Media was left alone for 72 hours, and then changed every 2 days until cell lysates were collected at 7 days. mRNA was extracted and qPCR was performed to probe for Fyn and S100β expression levels.

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 Rat - Schwann cells Fyn using ON-TARGETplus Rat Fyn siRNA from Dharmacon.

Manufacturer protocol

Download the product protocol from Dharmacon for ON-TARGETplus Rat Fyn siRNA below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing siRNA / miRNA gene silencing Rat - Schwann cells Fyn using ON-TARGETplus Rat Fyn siRNA from Dharmacon. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.