| Species : |
Human |
| Source : |
E.coli |
| Tag : |
His |
| Description : |
Pyruvate kinase catalyzes the conversion of phosphoenolpyruvate (PEP) to pyruvate, the last reaction of glycolysis. The M2 isozyme of pyruvate kinase, or PKM2 is the only isozyme expressed in cancer cells, and have been used clinically as tumor maker for detection of early cancer and monitor progress of cancer treatment. |
| Bio-activity : |
PKM2 activity was measured by an enzyme coupled assay utilizing lactic-acid dehydrogenase (LDH). PKM2 activity is reflected in the conversion of nicotinamide adenine dinucleotide acid (NADH) to nicotinamide adenine dinucleotide (NAD+). The amount of NADH is measured by colorimetric absorbance at 340 nm. The concentration of NADH can be calculated according to its extinction coefficient (6220/ M-1cm-1 at 340 nm). The purified PKM2 was activated by fructose 1,6-bisphosphate (FBP). The presence of FBP lowered the Km for substrate PEP by 20 fold. |
| AA Sequence : |
MGSSHHHHHHSSGLVPRGSHMSKPHSEAGTVRTATESFASDPILYRPVAVALDTKGPEIRGSLGSKKGVNLPGAAVDLPAVSEKDIQDLKKMMIGRCNRAGKPVICATQMLESMIKKPRPFKCCSGAIIVLTKSGRSAHQVARYRPRAPIPAFIQTQQLHAAMADTFLEHMCRLDIDSPPITARNTGIICTIGPASRSVETLKEMIKSGMNVARLNFSHGTHEYHAETIKNTGLIKGSGTAEVELKKGATLKITLDNAYMEKCDENILWLDYKNICKVVEVGSKIYVDDGLISLQVKQKGADFLVTEVENGFGVEQDVDMVFASFIRKASDVHEVRKVLGEKGKNIKIISKIENHEGVRRFDEILEASDGIMVARGDLGIEIPAEKVFLAQTRAEGSDVANAVLDGADCIMLSGETAKGDYPLEAVRMQHLIAREAEAAIYHLQLFEELRRLAPITSDPTEATAVGAVEASIAVTRNPQTARQAHLYRGIFPVLCKDPVQEAWAEDVDLRVNFAMNVGKARGFFKKGDVVIVLTGWRPGSGFTNTMRVVPV |
| Purity : |
> 98% by SDS-PAGE |
| Usage : |
Useful for studying enzyme kinetics, screening for PKM2 modulators including activators and inhibitors, and selectivity profiling. Protein binding assay, antigen, ELSA, Western blot, crystallization, co-crystallization and crystal structure determination study. |
| Storage : |
Upon delivery aliquot and store at -80 centigrade. Avoid multiple freeze-thaw cycles. The protein is stable for 12 months at -80 centigrade, for 2-4 weeks at 4 centigrade. |
| Concentration : |
0.5 mg/mL |
| Storage Buffer : |
PKM2 with 2 mM DTT, 20% glycerol, 0.02% NaN3, 150 mM NaCl, 0.1 mM EDTA, 50 mM Tris-HCl pH7.5. |
