X-tremeGENE™ HP DNA Transfection Reagent

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Do not aliquot X-tremeGENE HP DNA Transfection Reagent; store in the original glass vials.

Upstream tips
Do not aliquot X-tremeGENE HP DNA Transfection Reagent; store in the original glass vials.

Publication protocol

HAoSMCs were transfected with 2 μg DNA encoding constitutively active SGK1S422D, inactive SGK1K127N in pcDNA3.1 vector, or empty vector as control (18) using X-tremeGENE HP DNA transfection reagent (Roche Applied Science) according to the manufacturer’s protocol. The cells were used 24 hours (confocal microscopy), 48 hours (RT-qPCR, luciferase assay, Western blotting), or 7 days (ALPL activity) after transfection. HAoSMCs were transfected with 10 nM SGK1 siRNA (s740, Thermo Fisher Scientific), 10 nM IKKα siRNA (s3076, Thermo Fisher Scientific), or 10 nM IKKβ siRNA (s223928, Thermo Fisher Scientific) or with 10 nM negative control siRNA (4390843, Thermo Fisher Scientific) using siPORT amine transfection agent (Thermo Fisher Scientific) according to the manufacturer’s protocol. The cells were used 48 hours (RT-qPCR), 7 days (ALPL activity), or 14 days (calcium deposition) after silencing. Transfection and silencing efficiency were determined by real-time quantitative PCR (RT-qPCR).



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Manufacturer protocol

Download the product protocol from Sigma-Aldrich for X-tremeGENE™ HP DNA Transfection Reagent below.

Download manufacturer protocol

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