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Encompassing Amino Acids: full length
Applications: Useful for studying enzyme kinetics, screening inhibitors, and selectivity profiling.
Assay Conditions: Varying amounts of enzyme were incubated in 5 mM MOPS, pH 7.2, 2.5 mM β-glycerophosphate, 4 mM MgCl2, 2.5 mM MnCl2, 1 mM EGTA, 0.4 mM EDTA, 0.01 mM DTT, 0.05 mM ATP, 10 µg of S6K synthetic peptide substrate, and 32 µCi of [33P]-ATP was incubated at 30°C for 15 minutes. Then spot reaction on phosphocellulose paper, fix in 1% phosphoric acid, and assay with a scintillation counter.
Background: PIM1 is a proto-oncogene that belongs to a family of serine/threonine protein kinases that are highly conserved through evolution in multicellular organisms. Originally identified from Moloney murine leukemia virus induced T-cell lymphomas in mice, PIM1 is involved in the control of cytokine-mediated cell proliferation, differentiation and survival of lymphoid and myeloid cells as well as others (1). Expression of PIM1 can be stimulated by a variety of growth factors and is regulated at four different levels: transcriptional, post-transcriptional, translational and post-translational (2). Expression of PIM1 is mediated through activation of the JAK/STAT pathway.
Description: Recombinant human serine/threonine-protein kinase PIM1 (PIM1), GenBank Accession No. NM_002648, full-length with an N-terminalGST tag and expressed in a baculovirus infected Sf9 insect cell expression system. MW = 62 kDa.
Format: Aqueous buffer solution.
Formulation: 50 mM Tris-HCl, pH 7.5, 150mM NaCl, 10 mM glutathione, 0.1 mM EDTA,0.1 mM PMSF, 0.25 mM DTT, 25% glycerol
Genbank: NM_002648
Host Cell Line: Sf9 cells
Purity: ≥75%
Storage Stability: At least 6 months at -80°C.
Tags: N-terminal GST tag
Uniprot: P11309
Warnings: Avoidfreeze/thaw cycles. Storing diluted enzyme isnot recommended, if necessary, use carrierprotein (BSA 0.1 - 0.5%).
Biosafety Level: Not applicable (BSL-1)
References: 1. Meeker, T.C., et al., J. Cell. Biochem. 1987; 35(2): 105-112.
2. Friedmann, M. et al., Arch. Biochem Biophys. 1992; 298(2): 594-601.