PCSK9 siRNA (h)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	The A549 human LAD cell line was obtained from the American Type Culture Collection (Manassas, VA, USA). Cells were routinely grown in RPMI-1640 medium (Hyclone; GE Healthcare, Chalfont, UK) containing 10% fetal bovine serum (FBS; Hyclone), 100 U/ml penicillin (Sigma-Aldrich, Merck-Millipore, Darmstadt, Germany) and 100 µg/ml streptomycin (Sigma-Aldrich) at 37°C in a humidified atmosphere containing 5% CO2. The medium was replaced every 2–3 days and upon reaching 80% confluence, they were passaged at a 1:2 ratio. A549 cells were transfected with 100 nM PCSK9 siRNA (cat. no. sc-45482; Santa Cruz Biotechnology, Inc., Dallas, TX, USA; Genbank ID for PCSK9: NM_174936) or control siRNA (scrambled siRNA, a universal negative control; cat. no. sc-37007; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) with GeneSilencer siRNA transfection reagent (Genlantis, San Diego, CA, USA), according to the manufacturer's instructions. At 48 h after transfection, the efficiency of siRNA-mediated PCSK9 knockdown was determined by western blot analysis.

Protocol tips

The A549 human LAD cell line was obtained from the American Type Culture Collection (Manassas, VA, USA). Cells were routinely grown in RPMI-1640 medium (Hyclone; GE Healthcare, Chalfont, UK) containing 10% fetal bovine serum (FBS; Hyclone), 100 U/ml penicillin (Sigma-Aldrich, Merck-Millipore, Darmstadt, Germany) and 100 µg/ml streptomycin (Sigma-Aldrich) at 37°C in a humidified atmosphere containing 5% CO2. The medium was replaced every 2–3 days and upon reaching 80% confluence, they were passaged at a 1:2 ratio. A549 cells were transfected with 100 nM PCSK9 siRNA (cat. no. sc-45482; Santa Cruz Biotechnology, Inc., Dallas, TX, USA; Genbank ID for PCSK9: NM_174936) or control siRNA (scrambled siRNA, a universal negative control; cat. no. sc-37007; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) with GeneSilencer siRNA transfection reagent (Genlantis, San Diego, CA, USA), according to the manufacturer's instructions. At 48 h after transfection, the efficiency of siRNA-mediated PCSK9 knockdown was determined by western blot analysis.

Publication protocol

The A549 human LAD cell line was obtained from the American Type Culture Collection (Manassas, VA, USA). Cells were routinely grown in RPMI-1640 medium (Hyclone; GE Healthcare, Chalfont, UK) containing 10% fetal bovine serum (FBS; Hyclone), 100 U/ml penicillin (Sigma-Aldrich, Merck-Millipore, Darmstadt, Germany) and 100 µg/ml streptomycin (Sigma-Aldrich) at 37°C in a humidified atmosphere containing 5% CO2. The medium was replaced every 2–3 days and upon reaching 80% confluence, they were passaged at a 1:2 ratio. A549 cells were transfected with 100 nM PCSK9 siRNA (cat. no. sc-45482; Santa Cruz Biotechnology, Inc., Dallas, TX, USA; Genbank ID for PCSK9: NM_174936) or control siRNA (scrambled siRNA, a universal negative control; cat. no. sc-37007; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) with GeneSilencer siRNA transfection reagent (Genlantis, San Diego, CA, USA), according to the manufacturer's instructions. At 48 h after transfection, the efficiency of siRNA-mediated PCSK9 knockdown was determined by western blot analysis.

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

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing siRNA / miRNA gene silencing Human - A549 PCSK9 using PCSK9 siRNA (h) from Santa Cruz Biotechnology.

Manufacturer protocol

Download the product protocol from Santa Cruz Biotechnology for PCSK9 siRNA (h) below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing siRNA / miRNA gene silencing Human - A549 PCSK9 using PCSK9 siRNA (h) from Santa Cruz Biotechnology. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.