PLK4, GST-tag Recombinant

Catalog #

40236

$410 *

Size: 10 µg

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Description

Recombinant human PLK4 (polo like kinase 4), encompassing amino acids 1-836. This construct contains an N-terminal GST-tag. The recombinant protein was affinity purified and is kinase active.

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Synonyms

polo-like kinase 4, serine/threonine protein kinase 18 (STK18), SAK

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Product Data Gallery

4-20% SDS-PAGE Coomassie Staining

Specific Activity

Product Info

Storage and Usage

Citations

Species

Human

Construct

PLK4 (GST-1-836)

Host Species/Expression System

Sf9 Insect Cells

Purity

≥70%

Format

Aqueous buffer solution

Formulation

50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 10 mM Glutathione, 0.1 mM EDTA, 0.25 mM DTT, 0.1 mM PMSF, 25% glycerol

140 kDa

Amino Acids

1-836

Specific Activity

1.8 pmol/min/μg

Genbank #

NM_014264

UniProt #

O00444

Tag(s)

N-terminal GST-tag

References

1. Kleylein-Sohn, J. et al., Dev. Cell. 2007; 13: 190-202.
2. Ko, M.A. et al., Nature Genet. 2005; 37: 883-888.

Storage/Stability

At least 6 months at –80°C.

Assay Conditions

PLK4 activity was measured by using myelin basic protein (MBP) substrate diluted in 100 mM MOPS buffer (pH 6.5) to a final concentration of 0.5 mg/ml in the ADP Glo™ assay (Promega #V9101). Reaction was initiated by mixing increasing amounts of the PLK4 with 25 µM ATP in 40 mM Tris-HCl, pH 7.4, 20 mM MgCl₂, 0.1 mg/ml BSA prepared with 250 µM DTT and substrate at final concentration of 25 µg/ml substrate.

After a 40-minute incubation at 37°C, the reaction was terminated by addition of the AMP-Glo™ Reagent followed by a subsequent 40-minute incubation at room temperature. Kinase Detection Reagent was then added and incubated for another 30 minutes. Detection of luminescence was measured using the Luminescence Module Protocol on GloMax®-Multi Microplate reader. The corrected activity (RLU) was calculated by removing the blank value for each sample divided by the (specific activity of ADP in RLU/pmol)*(Reaction time in min)*(Enzyme amount in µg or mg). The blank was determined from a “no kinase” sample by replacing the enzyme working solution with an equal volume of Kinase Dilution Buffer IX (1X).

Instructions for Use

Thaw on ice and gently mix prior to use. DO NOT VORTEX. Perform a quick spin before opening. Aliquot into small volumes and flash freeze for long term storage. Avoid multiple freeze/thaw cycles.

Applications

Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.

Shipping Temperature

-80°C

Warnings

Avoid freeze/thaw cycles.

Scientific Category

Kinase/Serine-Threonine

Regulatory Status

Research Use Only

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Datasheets/Protocols

40236 Lot 230210

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