HEK293 cells were transfected with 5 μg of (Myc-DDK-tagged)-Human CD59 plasmid (OriGene) and 2.5 μl of EndoFectin Plus transfection reagent (GeneCopoeia) in a confluent 35-mm plate according to the manufacturer's protocol. After transfection for 72 hours, the HEK293 cells were lysed with immunoprecipitation (IP) lysis buffer (Thermo Scientific). The expression of Myc-DDK–tagged human CD59 was confirmed by detecting DDK with anti-DDK monoclonal antibody (OriGene) by Western blotting.
Endofectin plus transfection reagent
Manufactured by GeneCopoeia
Sourced in China, United States
EndoFectin Plus is a transfection reagent designed for efficient delivery of nucleic acids into a wide range of cell lines. It facilitates the uptake of DNA, RNA, and other macromolecules into the target cells.
Automatically generated - may contain errors
Lab products found in correlation
Myc ddk tagged human cd59 plasmid, by OriGene (2 mentions)
Anti ddk monoclonal antibody, by OriGene (2 mentions)
Ip lysis buffer, by Thermo Fisher Scientific (2 mentions)
6 cm culture dishes, by Corning (1 mentions)
Mission lightswitch luciferase assay reagent, by Merck Group (1 mentions)
Secrete pair dual luminescence assay kit, by GeneCopoeia (1 mentions)
Fluostar omega microplate reader, by BMG Labtech (1 mentions)
4 protocols using endofectin plus transfection reagent
1
Overexpression of Human CD59 in HEK293 Cells
2
NF-κB Signaling Pathway Modulation
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HUVECs were seeded into 6cm culture dishes (Corning, Lowell, MA) for overnight. Subsequently, 1μg of pNF-κB-M98 (pReceiver-M98, Funeng, Guangdong, China) or pcDNA was transfected into HUVECs using Endofectin-Plus transfection Reagent (Genecopoeia, Guangdong, China), according to the manufacturer's instruction. After transfection for 43h, cells were pretreated with or without DT-13 at 1μM for 1h prior to TNF-α (10 ng/mL) induction for 4h. The cells were collected for Western blot analysis.
3
Luciferase Assay for 3'-UTR Analysis
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For luciferase reporter assay, MISSION 3′-UTR Lenti GoClone HOXB9 (HUTR10238) and appropriate negative control (HUTR001C) from SwitchGear Genomics (Carlsbad, CA, USA) were used and the viral particles were added at MOI (multiplicity of infection) = 2.5. In case of EREG, 1 µg of pEZX-MT05 vector containing UTR for EREG (HmiT004978) or appropriate control vector (CmiT000001-MT05) were transfected into DLD-1 cells using EndoFectin Plus Transfection Reagent (GeneCopoeia, Rockville, MD, USA). The luciferase activity was measured using the MISSION LightSwitch Luciferase Assay Reagent (Sigma Aldrich) or SecretePair Dual Luminescence assay kit (GeneCopoeia), respectively, using FLUOstar Omega Microplate reader (BMG Labtech, Ortenberg, Germany).
4
Overexpression of Human CD59 in HEK293 Cells
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