Code | CSB-EP017613HU |
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Size | $224 |
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This Human PCK1 recombinant protein was produced in E.coli, where the gene sequence encoding Human PCK1 (1-622aa) was expressed with the N-terminal 6xHis tag. The purity of this PCK1 protein was greater than 90% by SDS-PAGE.
PCK1 plays a critical role in the gluconeogenesis pathway. This pathway helps maintain blood sugar levels when they are low due to fasting or extended periods without eating. It allows liver and kidney cells to synthesize glucose from non-sugar substrates to supply to other tissues, including the brain. The activity of PCK1 is regulated by various factors, including hormones such as insulin and adrenaline, energy status, and glucose concentration. Insulin inhibits the activity of PCK1, promoting glucose uptake and storage, while stress hormones like adrenaline can stimulate PCK1 activity, increasing glucose release. PCK1 is crucial for maintaining normal blood sugar levels and energy balance. Defects or abnormal activity of PCK1 are associated with diabetes, obesity, and metabolic disorders. Due to its importance in glucose metabolism, PCK1 has become a potential drug target. Some research is exploring the possibility of regulating PCK1 activity to treat diseases related to blood sugar regulation.
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I wanted to mention a critical thing about this protein. It needs Manganese (Mn2+) to fold properly. We found that our in house protein was behaving strangely in the presence of inhibitors and this was due to the presence of aberrantly folded protein. We had prepared His tagged material and to purify it metal affinity chromatography was used which depletes the protein from any Mn2+. I would highly recommend adding MnCl2 (1-2mM) to the final product preferably immediately after purification. Particularly if you are using metal affinity resin (talon or some other). Even if you are not I would strongly suggest inclusion of Manganese in the final buffer prior to freezing and shipment.
And:
One other point. The enzyme has a very active cysteine residue in the active site that is required for catalysis. It needs to be reduced. So use DDT (2-5 mM) or TCEP (1mM) in the final buffer.
Please let me know if adding MnCl2 and DDT or TCEP to the final product will be possible.