AllStars Hs Cell Death siRNA

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Add diluted siRNA to diluted Lipofectamine® RNAiMAX Reagent (1:1 ratio).  Incubate for 5 minutes at room temperature and add to cells.  Incubate cells for 3 days at 37°C.

Protocol tips
Add diluted siRNA to diluted Lipofectamine® RNAiMAX Reagent (1:1 ratio).  Incubate for 5 minutes at room temperature and add to cells.  Incubate cells for 3 days at 37°C.

Publication protocol

Cells were transfected with Sensor siRNAs at the indicated concentrations by using Lipofectamine RNAiMAX (Invitrogen) following the manufacture’s instruction. AllStars Negative Control siRNA and AllStars Human Cell Death Control siRNA (Qiagen) were transfected as controls. To validate the Sensor siRNAs and to test knockdown efficiency, U2OS cells were transfected with Sensor siRNAs at the indicated concentrations. Cells were collected 72 hours post-transfection for total cell lysate preparation or total RNA isolation using RNeasy Mini Kit (Qiagen) followed by cDNA conversion using High Capacity cDNA Reverse Transcription Kit (Applied Biosystems). To measure mRNA level, quantitative real-time PCR was performed using SYBR Green method on a 7900HT Fast Real-Time PCR System platform (Applied Biosystems). GAPDH mRNA was measured as an endogenous control. Each quantitative RT-PCR assay was performed in triplicate. To measure protein level, total cell lysates were subjected to immunoblotting as previously described (37).

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Manufacturer protocol

Download the product protocol from Qiagen for AllStars Hs Cell Death siRNA below.

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