IMP-6462

LATS1

Recombinant LATS1 protein was expressed in a baculovirus expression system as the full length human protein (accession number NP_004681.1) with an N-terminal FLAG tag. The molecular weight of the protein is 128.1 kDa.

LATS1 (Large Tumor Suppressor Kinase 1) is a putative serine/threonine kinase that localizes to the mitotic apparatus and complexes with cell cycle controller CDC2 kinase in early mitosis. It is phosphorylated in a cell-cycle dependent manner, with late prophase phosphorylation remaining through metaphase. The N-terminal region of LATS1 binds CDC2 to form a complex showing reduced H1 histone kinase activity, indicating a role as a negative regulator of CDC2/cyclin A. In addition, the C-terminal of it domain binds to its own N-terminal region, suggesting potential negative regulation through interference with complex formation via intramolecular binding. Biochemical and genetic data suggest that LATS1 play a significant role as a tumor suppressor which is supported by studies in knockout mice showing development of soft-tissue sarcomas, ovarian stromal cell tumors and a high sensitivity to carcinogenic treatments.

Insect cells

Human

FLAG

Recombinant LATS1 protein is supplied in 25 mM HEPES-NaOH pH 7.5, 300 mM NaCl, 10% glycerol, 0.04% Triton X-100, and 0.5 mM TCEP.

128.1 kDa

>73%

Enzyme kinetics, inhibitor screening, and selectivity profiling

Recombinant proteins in solution are temperature sensitive and must be stored at -80 centigrade to prevent degradation. Avoid repeated freeze/thaw cycles and keep on ice when not in storage.

Recombinant LATS1 protein gel 10% SDS-PAGE gel with Coomassie blue staining MW: 128.1 kDa Purity: >73%

HTRF assay for LATS1 protein activity 1 μM STK S1 substrate was incubated with different concentrations of LATS1 protein in a 10 μL reaction system containing 1×Enzymatic Buffer, 5 mM MgCl2, 1 mM DTT and 100 uM ATP for 1 hr. Then 10 μL detection reagents containing TK antibody and SA-XL665 (each of which was 1:100 diluted with 1× Detection Buffer) were added and incubated with the reactions for 30 min. All the operations and reactions were performed at room temperature.HTRF KinEASE STK assay was used to detect the enzymatic activity.

HTRF assay for LATS1 protein activity 1 μM STK S1 substrate was incubated with different concentrations of LATS1 protein in a 10 μL reaction system containing 1×Enzymatic Buffer, 5 mM MgCl2, 1 mM DTT and 100 uM ATP for 1 hr. Then 10 μL detection reagents containing TK antibody and SA-XL665 (each of which was 1:100 diluted with 1× Detection Buffer) were added and incubated with the reactions for 30 min. All the operations and reactions were performed at room temperature.HTRF KinEASE STK assay was used to detect the enzymatic activity.