GeneSilencer® Transfection Reagent

Protocol tips

Upstream tips	Protocol tips	Downstream tips
This Kit is compatibile with diverse growth conditions (with and without serum).	The ratio of siRNA (micrograms) to Gene Silencer (microliters) was 1:5. Incubate at room temp for 5-30 min.

Upstream tips
This Kit is compatibile with diverse growth conditions (with and without serum).
Protocol tips
The ratio of siRNA (micrograms) to Gene Silencer (microliters) was 1:5. Incubate at room temp for 5-30 min.

Publication protocol

AllStars negative siRNA control, Pgrmc1 siRNA duplexes (5′-TTGGATAATACTCTTGGTTAA-3′), and ERK5 siRNA duplexes (5′-GGCCATCAAGAAGATACCTAA-3′) were purchased from QIAGEN. Briefly, C6 cells were transfected with the siRNA duplexes using Gene Silencer reagent (Genlantis, San Diego, CA) per manufacturer's protocol. The ratio of siRNA (micrograms) to Gene Silencer (microliters) was 1:5. RNA was isolated 24 h after transfection. Silencing of Pgrmc1 and ERK5 expression was assessed by real-time PCR. To assess the roles of Pgrmc1 and ERK5 in mediating P4-induced BDNF release, cells were plated in anti-BDNF antibody-coated Nunc MaxiSorp immunoplates, as described in BDNF ELISA in situ. Twenty-four hours after plating, cells were transfected with siRNA duplexes and returned to a CO2 incubator for another 24 h before treatment with 1 nm P4 or P4-BSA. BDNF released into the media was measured by ELISA in situ after 18 h.

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Manufacturer protocol

Download the product protocol from Genlantis for GeneSilencer® Transfection Reagent below.

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