ON-TARGETplus Rat Atf4 (79255) siRNA - SMARTpool

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Gene silencing was performed using siRNAs. GRP78-specific siRNAs were synthesized by Shanghai GenePharma (Shanghai, China) with the sense strand sequence of 5′-GAGGCGUAUUUGGGAAAGATT-3′.The scrambled siRNA has the sense sequence of 5′-UUCUCCGAACGUGUCACGUTT-3′. To efficiently knock down ATF4, we used a pool of four different siRNAs (ON-TARGET plus SMART pool Rat ATF4; Thermo Scientific Dharmacon) to target rat ATF4 mRNA. Control cultures were incubated with non-targeting siRNAs (ON-TARGET plus non-targeting siRNA#1; Thermo Scientific Dharmacon).Transfection of siRNAs was carried out using Lipofectamine™ 2000 (Invitrogen, Grand Island, NY, USA), according to the manufacturer's instructions. Briefly, siRNA and Lipofectamine™ 2000 reagent were mixed in Opti-MEM medium (Invitrogen) and incubated for 30 min at room temperature to allow the complex formation. Cells were washed with Opti-MEM medium, and the transfection mixture was added. 6 h after transfection, cells were washed and cultured for 24 h in complete medium containing 10% FCS and 5% FBS. The silencing efficacy was evaluated by western blotting.

Protocol tips

Gene silencing was performed using siRNAs. GRP78-specific siRNAs were synthesized by Shanghai GenePharma (Shanghai, China) with the sense strand sequence of 5′-GAGGCGUAUUUGGGAAAGATT-3′.The scrambled siRNA has the sense sequence of 5′-UUCUCCGAACGUGUCACGUTT-3′. To efficiently knock down ATF4, we used a pool of four different siRNAs (ON-TARGET plus SMART pool Rat ATF4; Thermo Scientific Dharmacon) to target rat ATF4 mRNA. Control cultures were incubated with non-targeting siRNAs (ON-TARGET plus non-targeting siRNA#1; Thermo Scientific Dharmacon).Transfection of siRNAs was carried out using Lipofectamine™ 2000 (Invitrogen, Grand Island, NY, USA), according to the manufacturer's instructions. Briefly, siRNA and Lipofectamine™ 2000 reagent were mixed in Opti-MEM medium (Invitrogen) and incubated for 30 min at room temperature to allow the complex formation. Cells were washed with Opti-MEM medium, and the transfection mixture was added. 6 h after transfection, cells were washed and cultured for 24 h in complete medium containing 10% FCS and 5% FBS. The silencing efficacy was evaluated by western blotting.

Publication protocol

Gene silencing was performed using siRNAs. GRP78-specific siRNAs were synthesized by Shanghai GenePharma (Shanghai, China) with the sense strand sequence of 5′-GAGGCGUAUUUGGGAAAGATT-3′.The scrambled siRNA has the sense sequence of 5′-UUCUCCGAACGUGUCACGUTT-3′. To efficiently knock down ATF4, we used a pool of four different siRNAs (ON-TARGET plus SMART pool Rat ATF4; Thermo Scientific Dharmacon) to target rat ATF4 mRNA. Control cultures were incubated with non-targeting siRNAs (ON-TARGET plus non-targeting siRNA#1; Thermo Scientific Dharmacon).Transfection of siRNAs was carried out using Lipofectamine™ 2000 (Invitrogen, Grand Island, NY, USA), according to the manufacturer's instructions. Briefly, siRNA and Lipofectamine™ 2000 reagent were mixed in Opti-MEM medium (Invitrogen) and incubated for 30 min at room temperature to allow the complex formation. Cells were washed with Opti-MEM medium, and the transfection mixture was added. 6 h after transfection, cells were washed and cultured for 24 h in complete medium containing 10% FCS and 5% FBS. The silencing efficacy was evaluated by western blotting.

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 - PC12 Atf4 using ON-TARGETplus Rat Atf4 (79255) siRNA - SMARTpool from Horizon Discovery Ltd..

Manufacturer protocol

Download the product protocol from Horizon Discovery Ltd. for ON-TARGETplus Rat Atf4 (79255) siRNA - SMARTpool below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing siRNA / miRNA gene silencing Rat - PC12 Atf4 using ON-TARGETplus Rat Atf4 (79255) siRNA - SMARTpool from Horizon Discovery Ltd.. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.