TAK1_TAB1 complex [6His-tagged]

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Species Reactivity: Human  

Source: Sf21 insect cell-baculovirus ex ­pression  

Formula: 50 mM Tris/HCl pH 7.5, 0.1 mM EGTA, 150 mM NaCl, 270 mM sucrose, 0.03% Brij, 0.1% b-Mercapto-ethanol, 1 mM Benzamidine, 0.2 mM PMSF  

Concentration: 1 mg/ml  

Molecular Weight: ~44.5 kDa  

Protein Sequence: Accession number: TAK1: NP_003179.1; TAB1: NP_006107.1. For full protein sequence information download the Certificate of Analysis pdf.  

Quality Control Protein Identification: Confirmed by mass spectrometry.  

Quality Control Activity: Activity Assay:  The specific activity of His-TAK1-TAB1 was determined using the method described by Hastie et al. (2006) with the enzyme being assayed at several concentrations.   His-TAK1-TAB1 was incubated for 10 minutes at 30 °C in kinase reaction buffer in the presence of EP3151 peptide substrate (300 µM) and [γ-32P]ATP (100 µM). Duplicate reactions were stopped by spotting the assay mixture onto Whatman P81 paper - capturing the phosphorylated substrate.   The radioactivity incorporated was measured on a scintillation counter and the enzyme’s mean specific activity was calculated.  His-TAK1-TAB1 specific activity:92.5 Units/mg (92.5 Units/ml)1 Unit = 1 nmole of phosphate incorporated into the substrate in 1 minuteSubstrate: EP3151 (RLGRDKYKTLRQIRQ)