ON-TARGETplus Human FURIN (5045) siRNA - SMARTpool

siRNA / miRNA gene silencing Human - A253 Furin

Experiment
siRNA / miRNA gene silencing Human - A253 Furin
Product
ON-TARGETplus Human FURIN (5045) siRNA - SMARTpool from Dharmacon
Manufacturer
Dharmacon

Protocol tips

Protocol tips
Add diluted siRNA to diluted
Lipofectamine® RNAiMAX
Reagent (1:1 ratio).

Incubate cells for 1–3 days at 37°C.

Publication protocol

A253 and Detroit 562 (D562) are human head and neck squamous cell carcinoma cell lines, cultured in S-MEM with 10% FBS and 2 mM L-glutamine [20]. For all SiRNA experiments, cells were transfected by Lipofectamine® RNAiMAX Reagent (Invitrogen, Carlsbad, CA) using the manufacturer’s protocol. The following SiRNAs were purchased from Thermo Scientific (Lafayette, CO): ON-TARGETplus Non-targeting siRNA #1 (#D-001810-01-0005) for control siRNA, ON-TARGETplus PCSK7 siRNA (#J-005988-07-0005) for PC7, ON-TARGETplus FURIN siRNA-SMARTpool (#L-005882-00-0005) for Furin, and ON-TARGETplus PCSK6 siRNA (#J-005983-05-0002) for PACE4.

Cell lysates were harvested two days after siRNA transfection and subjected to Western blot analysis as described [33]. For CoCl2 and PPP treatments, the transfected or non-transfected cells were incubated with drugs overnight. The treatment time with MG-132 was 30 min. For CHX treatment, the cells were incubated for different times as labeled in the corresponding figures. Regarding VEGF-A and VEGF-C analysis, the cells were first transfected with SiRNA for 2 days and serum-free media were collected after 1-day incubation. The conditioned media were concentrated with Microcon centrifugal filters (#42407) from Millipore (Billerica, MA) and used for Western blot analysis. The following antibodies were used: HIF-1α (#610958) from BD Biosciences (San Jose, CA), VEGF-A (#sc-152), VEGF-C (#sc-1881) and IGF-1Rβ (#sc-713) from Santa Cruz Biotechnology (Santa Cruz, CA), and GAPDH (#ab8245) from Abcam (Cambridge, MA). The quantification of Western blot results was performed by using ImageJ developed by the National Institutes of Health.

Full paper   Login or join for free to view the full paper.

Reviews

ON-TARGETplus Human FURIN (5045) siRNA - SMARTpool from Dharmacon has not yet been reviewed for this experiment

We'd love it if you would be the first to write a review!

Discussion

Start your discussion

Share your thoughts or question with experts in your field

Start a discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing siRNA / miRNA gene silencing Human - A253 Furin using ON-TARGETplus Human FURIN (5045) siRNA - SMARTpool from Dharmacon.

View full paper   Login or join for free to view the full paper.

Manufacturer protocol

Download the product protocol from Dharmacon for ON-TARGETplus Human FURIN (5045) siRNA - SMARTpool below.

Download PDF Download manufacturer protocol

Videos

Check out videos that might be relevant for performing siRNA / miRNA gene silencing Human - A253 Furin using ON-TARGETplus Human FURIN (5045) siRNA - SMARTpool from Dharmacon. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.

We haven't found any additional videos for this experiment / product combination yet.

Outsource your experiment

Fill out your contact details and receive price quotes in your Inbox

  Outsource experiment
Become shareholder Discussions About us Contact Privacy Terms