Proper splicing of each intron for all IFN-λ2 construct was verified by PCR amplification across each splice sites using primers listed below. PCR products were run on a 2% agarose gel containing HydraGreen safe DNA dye (ACTGene) and imaged with a Syngene G:Box Chemi XT4 gel doc system.
G box chemi xt4 gel doc system
Manufactured by Syngene
Sourced in Italy
The G:Box Chemi XT4 is a gel documentation system designed for imaging and analyzing nucleic acid and protein gels. It features a high-resolution camera, automated image capture, and analysis software to accurately document and quantify gel-based experiments.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Horseradish peroxidase conjugated goat anti mouse igg secondary antibody, by Santa Cruz Biotechnology (1 mentions)
Polyvinylidene difluoride pvdf membrane, by Roche (1 mentions)
Sigmamarker, by Merck Group (1 mentions)
Anti beta actin antibody ab8226, by Abcam (1 mentions)
3 protocols using g box chemi xt4 gel doc system
1
Verification of IFN-λ2 Intron Splicing
2
FLAG Protein Expression in HEK293T Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Wild‐type and mutant (G306R) plasmids were transiently transfected into HEK293T (human embryonic kidney) cells cultured in Dulbecco's Modified Eagle's Medium (DMEM), using Lipofectamine 2000 (Invitrogen, Carlsbad, CA) according to the manufacturer's protocol. Total protein extracts (20 μg) from cell lysates were electrophoretically separated on 10% denaturing polyacrylamide gels and then transferred to Polyvinylidene difluoride (PVDF) membranes (Roche, Basilea, Switzerland). A high range SigmaMarker (Sigma‐Aldrich, Saint Louis, MO; molecular weight range, 36 000‐200 000 kDa) was used to characterize protein migration. Membranes were blocked in Tris‐buffered saline with 0.1% Tween 20 containing 5% non‐fat dried milk and reacted overnight at 4°C with the primary monoclonal antibody, anti‐FLAG mouse M2 clone (Sigma‐Aldrich) diluted 1:2000, and anti‐beta Actin antibody ab8226 (Abcam, Cambridge, UK), diluted 1:1000. After rinsing, filters were incubated with horseradish peroxidase‐conjugated goat anti‐mouse IgG secondary antibody (Santa Cruz), diluted 1:10000. Signals were developed using the enhanced chemiluminescent horseradish peroxidase (HRP) substrate Westar ETA C ultra (Cyanagen, Bologna, Italy), and specific protein bands were revealed using the G:BOX Chemi XT4 gel doc system (Syngene, Frederick, MD).
3
Verification of IFN-λ2 Intron Splicing
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Proper splicing of each intron for all IFN-λ2 construct was verified by PCR amplification across each splice sites using primers listed below. PCR products were run on a 2% agarose gel containing HydraGreen safe DNA dye (ACTGene) and imaged with a Syngene G:Box Chemi XT4 gel doc system.
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!