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TLK1B - Kinase Domain

Protein Name

Human TLK1b-kinase domain (Isoform 3)

Tag Name

 

Expression System

E. coli Rosetta-gami™ 2 (DE3)pLysS

Accession Number

 

Product Description

The TLK1B kinase domain (KD) comprises a coiled-coil region and the functional kinase domain spanning amino acids Ser227 to Leu530. Unlike the full-length TLK1 protein, the TLK1B KD lacks the N-terminal 226 amino acids and retains its catalytic activity. This domain is essential for its kinase activity.

Formulation

 

Specific Activity

 

Storage and Stability

20ºC for long term, 4ºC for short term use

Handling Recommendations

Proteins should be stored at cold temperatures when not in use.

To minimise freeze-thaw cycles, aliquot the protein into smaller portions of at least 10 μL to prevent evaporation and maintain consistent concentration.

Kinase Assay Protocol

1X Kinase reaction Buffer (KRB): 40 mM Tris-Cl (pH=7.5), 20 mM MgCl2, 2.5 % Glycerol, 0.1 mg/ml BSA
The desired concentration of TLK1B-KD (2X)- 2µM
Substrate dilution (2X): 2 µM in 1X KRB
ATP dilution (2X): 200 µM in 1X KRB

Assay Procedure:

1.The ADP-Glo kinase assay was performed in a solid, white, flat bottom 96-well plate.
2.The reaction volume of 20 µl with 2X 1 µM kinase TLK1B-KD, 2X 1 µM kinase substrate, and 2X 100 µM ATP in 1X Kinase Reaction Buffer (KRB).
3.The reaction was incubated for an hour at 30°C with gentle shaking.
4.20 µL of ADP-GLo reagent is added and incubated for 45 minutes.
5.40 µl of a Kinase Detection Reagent is added to each well and incubated for 60 minutes in the dark.
6.The readout was measured as luminescent values (Relative luminescent unit, RLU).

3. Stable Cell Lines

MDC1 Knockdown cell line (HeLa, SiHa, CasKi)
MDC1 Overexpressing cell line (HeLa, SiHa, CasKi)

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