Thermo Fisher 1-step human coupled in vitro transcription-translation kit (cat# 88881) was used to synthesize proteins expressed under the T7 promoter. To express the proteins, the protocol provided by the manufacturer was followed. In short, 5 μL of HeLa lysate was mixed with 2 μL reaction mixture and 1 μL accessory proteins. The reaction volume was then brought to 10 μL by adding 10 nM DNA plasmid and ultra-pure water. In cases where direct reconstitution of InterTag or InterCatch was desired, water was replaced by 5 mM 100% DOPC SUV solution so that the final concentration of SUV in the reaction was around 1 mM. The reactions were next transferred to a 384 conical well plate and incubated in the Synergy H1 plate reader (BioTek) at 30 °C for 3–4 h. The GFP, BFP, and sfCherry signals were monitored using plate readers at 400/450 nm, 488/528 nm, and 561/625 nm excitation/emission wavelengths, respectively. For experiments that required in vitro translation labeling system, a 1:50 dilution of FluoroTect™ GreenLys (Promega) was added to the reaction before incubation.
1 step human coupled in vitro transcription translation kit
Manufactured by Thermo Fisher Scientific
The 1-step human coupled in vitro transcription-translation kit is a laboratory tool designed to synthesize proteins from DNA templates. It provides a single-step process for transcribing and translating DNA into functional proteins without the need for separate steps. The kit includes the necessary reagents and components to enable this coupled in vitro process.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using 1 step human coupled in vitro transcription translation kit
1
In Vitro Protein Expression and Characterization
2
In vitro protein expression and labeling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Thermo Fisher 1-step human coupled in vitro transcription-translation kit (cat# 88881) was used to synthesize proteins expressed under the T7 promoter. To express the proteins, the protocol provided by the manufacturer was followed. In short, 5 µL of HeLa lysate was mixed with 2 µL reaction mixture and 1 µL accessory proteins. The reaction volume was then brought to 10 µL by adding 10 nM DNA plasmid and ultra-pure water. In cases where direct reconstitution of InterTag or InterCatch was desired, water was replaced by 5 mM 100% DOPC SUV solution so that the final concentration of SUV in the reaction was around 1 mM. The reactions were next transferred to a 384 conical well plate and incubated in the Synergy H1 plate reader (BioTek) at 30 °C for 3-4 h. The GFP, BFP, and sfCherry signals were monitored using plate readers at 400/450 nm, 488/528 nm, and 561/625 nm excitation/emission wavelengths, respectively. For experiments that required in vitro translation labeling system, a 1:50 dilution of FluoroTect™ GreenLys (Promega) was added to the reaction before incubation.
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!