| Products/Services Used | Details | Operation |
|---|---|---|
| Recombinant Proteins | The enzyme was incubated for 1 hour at 37°C in the following conditions: (Lane 1) 1 mM copper-1,10-phenanthroline, (Lane 2) 100 µM copper-1,10-phenanthroline, (Lane 3) 10 mM β-mercaptoethanol, (Lane 4) 1 mM β-mercaptoethanol. The gel was a 4-20% SDS-PAGE gel (GenScript, Piscataway, NJ, USA) run at 150V with Tris-MOPS buffer. Lane (L) is an AccuRuler RGB Pre-stained Protein Ladder (Biopioneer, San Diego, CA, USA).10 µL of IMAC eluate was diluted in buffer A containing 300 mM imidazole to yield a protein concentration of 1 mg/mL. 10 µL of this diluted Ni-NTA eluate was mixed with 5 µL of 4X SDS-PAGE loading dye containing 10 mM β-mercaptoethanol, vortexed, and heated to 96°C for 3 minutes. 10 µL of this solution was loaded onto a 4-20% PAGE gel from Genscript. | Get A Quote |
Isoprenoids are a remarkably diverse class of more than 25,000 biomolecules that include familiar compounds such as cholesterol, chlorophyll, vitamin A, ubiquinone, and natural rubber. The two essential building blocks of all isoprenoids, isopentenyl pyrophosphate (IPP) and dimethylallyl pyrophosphate (DMAPP), are ubiquitous in the three domains of life. In most eukaryotes and archaea, IPP and DMAPP are generated through the mevalonate pathway. We have identified two novel enzymes, mevalonate-3-kinase and mevalonate-3-phosphate-5-kinase from Thermoplasma acidophilum, which act sequentially in a putative alternate mevalonate pathway. We propose that mevalonate-3,5-bisphosphate is acted upon by a yet unidentified... More
