MdWRKY40 full length CDS inserted into the pHBT-AvrRpm 1 carrier and promoter segments of MdGLU into the pFRK1-LUC-nos carrier. Both plasmids were converted simultaneously from the protoplasm of apple callus and then expressed for 6 h at 24 °C. Subsequently, the protoplasm was suspended in 100 μL of cell lysate. The 5 μL cell extract and 20 μL 1 mmol·L− 1 4-MUG were incubated at 37 °C at 1 h, and the 100 μL of 0.2 mol·L− 1 sodium acetate was added to the termination reaction. The LUC activity was determined using the Luciferase Reporting Analysis System (Promega, Madison, WI, USA).
Luciferase reporting analysis system
Manufactured by Promega
Sourced in United States
The Luciferase reporting analysis system is a tool used to measure and quantify luciferase-based reporter gene expression in biological samples. It provides a sensitive and reliable method for monitoring gene expression, cellular activity, and other biological processes in a variety of experimental settings.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using luciferase reporting analysis system
1
Transient Expression of MdWRKY40 and MdGLU
Check if the same lab product or an alternative is used in the 5 most similar protocols
MdWRKY40 full length CDS inserted into the pHBT-AvrRpm 1 carrier and promoter segments of MdGLU into the pFRK1-LUC-nos carrier. Both plasmids were converted simultaneously from the protoplasm of apple callus and then expressed for 6 h at 24 °C. Subsequently, the protoplasm was suspended in 100 μL of cell lysate. The 5 μL cell extract and 20 μL 1 mmol·L− 1 4-MUG were incubated at 37 °C at 1 h, and the 100 μL of 0.2 mol·L− 1 sodium acetate was added to the termination reaction. The LUC activity was determined using the Luciferase Reporting Analysis System (Promega, Madison, WI, USA).
+ Expand
MdWRKY40 full length CDS inserted into the pHBT-AvrRpm 1 carrier and promoter segments of MdGLU into the pFRK1-LUC-nos carrier. Both plasmids were converted simultaneously from the protoplasm of apple callus and then expressed for 6 h at 24 °C. Subsequently, the protoplasm was suspended in 100 μL of cell lysate. The 5 μL cell extract and 20 μL 1 mmol·L− 1 4-MUG were incubated at 37 °C at 1 h, and the 100 μL of 0.2 mol·L− 1 sodium acetate was added to the termination reaction. The LUC activity was determined using the Luciferase Reporting Analysis System (Promega, Madison, WI, USA).
2
Investigating miR-218-5p regulation of TLR4
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
THP-1 cells were inoculated into 24-well plates for cell culture. In brief, a wild-type TLR4-3’UTR fragment containing the putative binding region of miR-218-5p or a mutated TLR4-3’UTR fragment containing the putative binding region of miR-218-5p was inserted into the luciferase reporter gene plasmid. Then, the above reporter plasmids and miR-218-5p mimic or miR-218-5p inhibitor were co-transfected into cells. After 48 h of transfection, luciferase activity was measured using the luciferase reporting analysis system (Promega, Madison, WI, USA) according to the manufacturer's instructions. The luciferase activities were normalized to Renilla fluorescence.
3
Validating miR-187-3p Binding to VPS9D1-AS1/FGFRL1
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The 3' untranslated region of wild type (wt) VPS9D1-AS1/FGFRL1 having binding sites with miR-187-3p was cloned to multi colony sites in the downstream of pmirGLO (Promega, USA) luciferase gene (VPS9D1-AS1-Wt/FGFRL1-Wt). At the same time, VPS9D1-AS1/FGFRL1 mutant (mut) vector was constructed (VPS9D1-AS1-Mut/FGFRL1-Mut). Afterward, VPS9D1-AS1-Wt/FGFRL1-Wt or VPS9D1-AS1-Mut/FGFRL1-Mut and miR-187-3p were co-transfected into PC-3 cells using Lipofectamine 2000. Forty-eight hour later, activities of firefly and renilla luciferase were assessed on the luciferase reporting analysis system (Promega, USA), and the renilla luciferase viability was utilized for standard processing on data.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!