TGFβR1, GST-Tag Recombinant

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Encompassing Amino Acids: 80-end

Applications: Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling. 

Assay Conditions: Kinase activity was measured using the ADP-Glo™ Kinase Assay Kit (Promega; Cat# V9101) which quantifies the amount of ADP produced. The ADP-Glo™ Reagent is added to terminate the reaction and deplete the remaining ATP. The Kinase Detection Reagent is then added to convert ADP to ATP and to measure the newly synthesized ATP using a luciferase reaction.TGFBR1 kinase activity was measured by using TGFBR1 peptide (KKKVLTQMSPSIRCS (pS)VS) diluted in 20 mM Tris-HCl, pH 7.5, to a final concentration of 1 mg/ml. Reaction was initiated by mixing increasing amounts of the TGFBR1 with 25 µM ATP in 40 mM Tris-HCl, pH 7.4, 20 mM MgCl₂, 0.1 mg/ml BSA prepared with 250 µM DTT and the 20 µg/ml substrate. After a 40-minute incubation at 37°C, the reaction was terminated by addition of the AMP-Glo™ Reagent followed by a subsequent 40-minute incubation at room temperature. Kinase Detection Reagent was then added and incubated for another 30 minutes. Detection of luminescence was measured using the Luminescence Module Protocol on GloMax®-Multi Microplate reader. The corrected activity (RLU) was calculated by removing the blank value for each sample divided by the (specific activity of ADP in RLU/pmol)*(Reaction time in min)*(Enzyme amount in µg or mg).  The blank was determined from a "no kinase" sample by replacing the enzyme working solution with an equal volume of Kinase Dilution Buffer IX (1X).

Description: Recombinant human TGFβR1 (transforming growth factor, beta receptor 1), also known as ALK5, encompassing amino acids 80-end. This construct contains an N-terminal GST tag. The recombinant protein was affinity purified and is kinase active.

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, 25% glycerol 

Genbank: BC071181

Host Cell Line: Sf9 cells

Purity: ≥90%

Storage Stability: At least 6 months at -80°C. 

Tags: N-terminal GST-tag

Uniprot: P36897

Warnings: Avoid freeze/thaw cycles.

Biosafety Level: Not applicable (BSL-1)

• Artikelnummer: BPS101575
• Hersteller: BPS Bioscience
• Hersteller Artikelnummer: 101575
• Green Labware: Nein
• Verpackungseinheit: 10 µg
• Mengeneinheit: STK
• Wirt: Insect