PIM2, GST-tag Recombinant

Catalog #

40153

$410 *

Size: 10 µg

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Description

Recombinant human full length PIM2. The constructs contain an N-terminal GST-tag. The protein was affinity purified and is active.

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Synonyms

PIM-2

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Product Data Gallery

4-20% SDS-PAGE Coomassie Staining

Specific Activity

Product Info

Storage and Usage

Citations

Species

Human

Construct

PIM2 (GST-Full Length)

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

61 kDa

Amino Acids

full length

Specific Activity

270 pmol/min/µg

Genbank #

NM_006875

UniProt #

Q9P1W9

Tag(s)

N-terminal GST-tag

Background

PIM2 is a serine threonine kinase that is present in all tissues, being most abundant in hematopoietic tissues, spleen, thymus, and peripheral blood leukocytes, as well as in testis, small intestine, and colon (1). It is highly expressed in human leukemic, lymphoma and colorectal adenocarcinoma cell lines. This suggests a role for PIM2 in proliferating cells as well as during meiosis. Similar to PIM1, PIM2 also acts as a pro-survival kinase and BAD protein is a legitimate PIM2 substrate (2).

References

1. Baytel, D. et al. Biochim. Biophys. Acta 1442: 274-285, 1998.
2. Yan, B. et al. J Biol Chem. 2003 Nov 14;278(46):45358-67.
3. Neri P, Bahlis NJ. Blood. 2013 Aug 29;122(9):1534-6.
4. Blanco-Aparicio C, Carnero A. Biochem Pharmacol. 2013 Mar 1;85(5):629-43.

Storage/Stability

At least 6 months at -80°C.

Assay Conditions

PIM2 activity was measured by using S6K Substrate (KRRRLASLR), 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 PIM2 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 Pim2 + 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 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

40153 Lot 240216

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