A20-specific small interfering RNA (siRNA) and non-targeting scrambled control siRNA were designed and purchased from Eurofins MWG Operon (siMAX siRNA). siRNAs were transfected into freshly isolated primary CD11b+ murine macrophages with the Amaxa Nucleofector device set to program Y001 using the Mouse Macrophage Nucleofector Kit. Cells were washed once 1 h after transfection and cultured for an additional 2 days before adiponectin and LPS stimulation.
Simax sirna
Manufactured by Eurofins
Sourced in Germany
SiMAX siRNA is a laboratory tool used for gene silencing experiments. It provides a reliable and efficient platform for the delivery of small interfering RNA (siRNA) into cells to knockdown target gene expression.
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Lab products found in correlation
3 protocols using simax sirna
1
Knockdown of A20 in Murine Macrophages
2
ESRP2 Knockdown in Huh-7 Cells
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The Huh-7 cell line (300156, CLS, Eppelheim, Germany) was cultured in Dulbecco’s Modified Eagle’s Media (DMEM) high glucose (D6429, Sigma Aldrich, St. Louis, MO, USA), supplemented with 10% FBS and 1% penicillin/streptomycin. Cells were cultured in a humidified chamber at a constant 37 °C and 5% O2. The knockdown of ESRP2 expression was performed using PepMute siRNA Transfection Reagent (SL100566, SignaGen Laboratories, Rockville, MD, USA). Cells were seeded at a density of 200.000 cells/well on a 6-well plate, where they reached approximate 50% confluence 24 h after seeding. Cells were transfected using the protocol provided by the manufacturer with 30 nM concentration of siRNA concentration. The cell medium was changed 24 h after transfection with complete medium, and cells were cultured for an additional 48 h before RNA extraction. siRNA sequences used for silencing were: siRNA-ESRP2 5′-GACUUAAUCCUCCUAGUUUtt-3′ [68 (link)] and siRNA-negative 5′-AGGUAGUGUAAUCGCCUUGtt-3′ (Non Specific Control 47% GC, Eurofins Genomics, Ebersberg, Germany) (siMAX siRNA, Eurofins Genomics, Ebersberg, Germany). Silencing was performed in three independent biological triplicates.
3
Keratin siRNA Knockdown in Colon Cancer
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HT-29 colon cancer cells were transfected for 8 h and overnight for 24 h with Pre-design Chimera RNAi (Abnova, Taipei, Taiwan) designed for KRT8 (H00003856-R01), KRT18 (H00003875-R01), and scrambled negative control Naito1 (R0017). Transfections of 30% confluent HT-29 cells were performed with 400 nM RNA interference in Lipofectamine2000 (Invitrogen, Carlsbad, CA) and diluted in Opti-MEM medium (Gibco). Cells were harvested after 72 h.
Caco-2 cells were transfected with siMAX siRNA (Eurofins Genomics, Ebersberg, Germany) designed for KRT8 (5′-GCCUCCUUCAUAGACAAGGUA(dTdT)-3′). The sequence for the K8 siRNA (clone ID TRCN0000062384) was obtained from the TRC Library Database, and nontarget (scr) siRNA (5′-CCUACAUCCCGAUCGAUGAUG(dTdT)-3′) was used as a control. Transfections of 20–30% confluent Caco-2 cells were performed with 50 pmol of siRNA in Lipofectamine2000 and diluted in Opti-MEM. Cells were harvested after 72 h.
Caco-2 cells were transfected with siMAX siRNA (Eurofins Genomics, Ebersberg, Germany) designed for KRT8 (5′-GCCUCCUUCAUAGACAAGGUA(dTdT)-3′). The sequence for the K8 siRNA (clone ID TRCN0000062384) was obtained from the TRC Library Database, and nontarget (scr) siRNA (5′-CCUACAUCCCGAUCGAUGAUG(dTdT)-3′) was used as a control. Transfections of 20–30% confluent Caco-2 cells were performed with 50 pmol of siRNA in Lipofectamine2000 and diluted in Opti-MEM. Cells were harvested after 72 h.
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