ON-TARGETplus Human PCSK7 (9159) siRNA - Individual

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Seed 5 × 10^5 cells	Final concentration of siRNAs-30 nM. Add siRNA with Xfect Reaction Buffer and vortex for 10 sec at high speed. Incubate for 10 min at room temperature and add to cells. Incubate cells at 37°C for 4 hr to overnight. Replace medium after 4h and incubate for another 48h.

Upstream tips
Seed 5 × 10^5 cells
Protocol tips
Final concentration of siRNAs-30 nM. Add siRNA with Xfect Reaction Buffer and vortex for 10 sec at high speed. Incubate for 10 min at room temperature and add to cells. Incubate cells at 37°C for 4 hr to overnight. Replace medium after 4h and incubate for another 48h.

Publication protocol

Expression plasmid of human IGFBP-7 was purchased from OriGene (#SC119176). Transient IGFBP-7 expression A253-5 cells (marked as “IGFBP-7”) were obtained using Xfect® transfection reagent (Clontech) according the manufacturer’s instructions. 5 × 105 of A253-5 cells were seeded in a 60 mm dish and cultured overnight. A total of 15 μg of plasmid was used for transfection procedure for each dish. A253-5 without transfection was labeled as a control (C) and cells transfected with pEGFP-N1 were marked as “Mock” in this study. For siRNA transfection, negative control siRNA and siRNA against IGFBP-7 (#s7241) were purchased from Invitrogen. Final concentration of both siRNAs at 30 nM was used in the transfection to obtain “IGFBP-7 + NC si” and IGFBP-7 + siRNA respectively.

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Manufacturer protocol

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