PIM3, Active, GST-tag Recombinant (Mouse)

Catalog #

41108

$410 *

Size: 10 µg

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Description

Recombinant mouse full length PIM3. The constructs contain an N-terminal GST-tag. The protein was affinity purified and is active.

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Synonyms

serine/threonine-protein kinase PIM3

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Product Data Gallery

4-20% SDS-PAGE Coomassie Staining

Specific Activity

Product Info

Storage and Usage

Citations

Species

Mouse

Construct

PIM3 (GST-Full Length) (Mouse)

Host Species/Expression System

Sf9 Insect Cells

Purity

≥90%

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, and 25% glycerol

63 kDa

Amino Acids

full-length

Specific Activity

785 pmol/min/µg

Genbank #

NM_145478

UniProt #

P58750

Tag(s)

N-terminal GST tag

References

1. Fujii, C. et al., Int. J. Cancer. 2005; 114:: 209-218.
2. Li, Y.-Y., et al., Cancer Res. 2006; 66: 6741-6747.

Storage/Stability

At least 6 months at –80°C.

Assay Conditions

PIM3 activity was measured by using RSK Substrate (KRRRLSSLRA), diluted in distilled water to a working concentration of 1 mg/ml, in a [33P]-ATP based assay. Reaction was initiated by mixing increasing amounts of PIM3 with 1250 pmoles of [33P]-ATP in 5 mM MOPS, pH 7.2, 2.5 mM β-glycerol-phosphate, 5 mM MgCl₂, 1 mM EGTA, 0.4 mM EDTA, 50 ng/µl BSA, 50 µM DTT and substrate at a final concentration of 200 µg/ml.

The reaction was initiated by addition of 5 µl of [33P]-ATP Assay Cocktail (50 µM of [33P]-ATP with 50 µM ATP) to 20 µl of Reaction Mix (10 µl of diluted PIM3 + 5 µl of Substrate Solution + 5 µl of distilled water), followed by a 15-minute incubation at 30°C. The reaction was terminated by spotting the reaction mixture on phosphocellulose P81 paper, air-dry and three 10-minute washes with 1% phosphoric acid solution. Radioactivity was measured in a scintillation counter. The corrected activity (RLU) was calculated by removing the blank value for each sample. The Kinase Specific Activity was calculated as follows: RLU / [(specific activity of [33P]-ATP in cpm/pmol)*(Reaction time in min)*(Enzyme amount in µg or mg)] * [(Reaction Volume) / (Spot Volume)]. The blank was determined from a “no substrate” sample by replacing the substrate solution with an equal volume of distilled water.

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 studying enzyme kinetics, screening inhibitors, and selectivity profiling.

Shipping Temperature

Dry ice

Warnings

Avoid freeze/thaw cycles

Scientific Category

Kinase - Serine/Threonine

Regulatory Status

Research Use Only

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Datasheets/Protocols

41108 Lot 240216

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PIM3, Active, GST-tag Recombinant (Mouse)

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