Jurkat cells (2.0 × 105) in 12-well plates were cotransfected with the expression vector pHβPr-1-neo encoding Tax, TaxΔC, TaxM22, Tax703, Tax(225-232) or Tax2B, together with pGK/β-gal and either κB-luc or CREluc using TransFectin (Bio-Rad Technologies) according to the manufacturer’s instructions. At 48 h after transfection the cell lysates were prepared, and the luciferase and β-galactosidase activities in the lysates were measured using both the Luciferase Assay System (Promega, Fitchburg, WI, USA) and the Galacto-Light System (Applied Biosystems, Foster City, CA, USA), respectively.
Galacto light system
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Galacto-Light System is a laboratory instrument used for the detection and quantification of galactose in various samples. It employs a bioluminescent assay method to measure galactose levels with high sensitivity and accuracy.
Automatically generated - may contain errors
3 protocols using galacto light system
1
Jurkat Cell Transcription Factor Assay
2
Transfection of Tax variants in Jurkat cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Jurkat cells (2.0 × 105) in 12-well plates were cotransfected with the expression vector pHβPr-1-neo encoding Tax, TaxΔC, TaxM22, Tax703, Tax(225-232) or Tax2B, together with pGK/β-gal and either κB-luc or CRE-luc using TransFectin (Bio-Rad Technologies) according to the manufacturer's instructions. At 48 h after transfection the cell lysates were prepared, and the luciferase and β-galactosidase activities in the lysates were measured using both the Luciferase Assay System (Promega, Fitchburg, WI, USA) and the Galacto-Light System (Applied Biosystems, Foster City, CA, USA), respectively.
3
Transient Transfection of HeLa Cells for β-Gal and Luc Reporter Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The plasmid pTN24, carrying the β-galactosidase (β-gal) and luciferase (luc) gene has been described elsewhere (31 (link)). HeLa cells have been transiently transfected with pTN24 and co-transfected with either empty pcDNA3 plasmid or full-length Jmjd6 in pcDNA3. β-gal and luc activity was detected in a luminometer (Lumat 9501, Berthold) 24 h post-transfection. Therefore, cells were lysed with passive lysis buffer of the dual-luciferase reporter assay system (Promega, Wisconsin, USA) for 15 min. Lysates were centrifuged at 14 000 g for 1 min. Luc activity was measured in 10 μl of the supernatant by using the dual-luciferase reporter assay system (Promega, Wisconsin, USA) and β-gal activity was analysed with the Galacto-Light system (Applied Biosystems) according to the manufacturer's instructions.
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!