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Encompassing Amino Acids: 1-346
Applications: Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Assay Conditions: 10 µl of varying concentrations of HPK1 was mixed with 5 µl of 1 mg/ml MBP protein and 5 µl of water. Reaction was initiated with 5 µl of 250 µM [33P]-ATP in 25 mM MOPS, pH 7.2, 12.5 mM β-glycerolphosphate, 25 mM MgCl2, 5 mM EGTA, 2 mM EDTA and 0.25 mM DTT and incubated at 30°C for 15 min. Reaction was terminated by spotting 20 µl of reaction solution onto pre-cut strips of P81 paper and reading in the presence of scintillation fluid and a scintillation counter.
Description: Human hematopoietic progenitor kinase 1 (HPK1) also known as mitogen activated protein kinase kinase kinase kinase 1 (MAP4K1), GenBank Accession No. NM_007181, a.a. 1-346, with N-terminal GST tag, MW = 65 kDa. HPK1 is expressed in Sf9 insect cells via a baculovirus expression system.
Format: Aqueous buffer solution
Formulation: 50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 10 mM glutatione, 0.1 mM EDTA, 0.25 mM DTT, 0.1 mM PMSF, 25% glycerol
Genbank: NM_007181
Host Cell Line: Sf9 cells
Storage Stability: At least 6 months at -80°C.
Tags: N-terminal GST tag
Uniprot: Q92918
Warnings: Avoidfreeze/thaw cycles. Storing diluted enzymeis not recommended, if necessary, usecarrier protein (BSA 0.1 - 0.5%).
Biosafety Level: Not applicable (BSL-1)
References: 1. Hu, M., et al., Genes Dev. 1996; 10: 2251-2264
2. Shui, J.R., et al., Nat. Immunol. 2007; 8: 84-91.