Related Experiment Video
Updated: Oct 18, 2025
06:09
Assaying the Kinase Activity of LRRK2 in vitro
Published on: January 18, 2012
22.7K
In vitro NLK Kinase Assay.
Sungho Moon1, Jiyoung Kim1, Eek-Hoon Jho1
1Department of Life Science, University of Seoul, Seoul, Korea.
Bio-Protocol
|October 1, 2021
Summary
This protocol details an in vitro kinase assay for nemo-like kinase and an efficient method for expressing and purifying Glutathione S-transferase (GST) fusion proteins using a mild lysis buffer.
Area of Science:
- Biochemistry
- Molecular Biology
- Protein Chemistry
Background:
- Nemo-like kinase (NLK) is a key regulator in various cellular processes.
- Characterizing NLK activity requires robust biochemical assays.
- Efficient production of recombinant proteins is crucial for biochemical studies.
Purpose of the Study:
- To provide a detailed protocol for an in vitro kinase assay specific to nemo-like kinase.
- To establish an efficient method for expressing and purifying Glutathione S-transferase (GST) fusion proteins.
Main Methods:
- Step-by-step instructions for performing an in vitro kinase assay for NLK.
- Utilization of a mild lysis buffer for protein expression and purification.
- Purification of GST-tagged fusion proteins.
Main Results:
- A reproducible protocol for NLK in vitro kinase assays was established.
- An efficient method for producing high-purity GST-NLK fusion proteins was developed.
- The protocol facilitates further biochemical and structural studies of NLK.
Conclusions:
- The presented protocol enables reliable in vitro kinase activity assessment of NLK.
- The described method for GST fusion protein production is efficient and scalable.
- This work provides valuable tools for researchers studying NLK function and regulation.
