| Cat. No.: | DHFR-375H |
| Product Overview: | Recombinant Dihydrofolate reductase protein, fused to His-tag at N-terminus, was expressed inE.coliand purified by using conventional chromatography techniques. MW = 23.6 kDa (207aa). |
| Description: | DHFR, also known as Dihydrofolate reductase, is anenzymethat reducesdihydrofolic acidtotetrahydrofolic acid, usingNADPHaselectron donor, which can be converted to the kinds of tetrahydrofolatecofactorsused in 1-carbon transfer chemistry. Dihydrofolate reductase deficiency has been linked to megaloblastic anemia. |
| Source: | E.coli. |
| Amino Acid Sequence: | MGSSHHHHHH SSGLVPRGSH MVGSLNCIVA VSQNMGIGKN GDLPWPPLRN EFRYFQRMTT TSSVEGKQNL VIMGKKTWFS IPEKNRPLKG RINLVLSREL KEPPQGAHFL SRSLDDALKL TEQPELANKV DMVWIVGGSS VYKEAMNHPG HLKLFVTRIM QDFESDTFFP EIDLEKYKLL PEYPGVLSDV QEEKGIKYKF EVYEKND |
| Form: | Liquid. In 20 mM Tris-HCl buffer (pH8.0) containing 0.1M NaCl 2mM DTT, and 30% glycerol. |
| Purity: | > 95% by SDS – PAGE. |
| Concentration: | 1 mg/ml (determined by Bradford assay). |
| Biological activity: | Specific activity is 1.5 - 2.5 units/ml and was obtained by measuring the oxidation of NADPH in absorbance at 340 nm during reaction. One unit will convert 1.0 umole of 7,8 dihydrofolate and beta-NADPH to 5,6,7,8-tetrahydrofolate and beta-NADP per minute at pH 6.5 at 25℃. |
| Activity Assay: | 1.Prepare a 3.2 ml reaction mixture into a suitable container: The final concentrations are 50 mM potassium phosphate, 0.72 mM DHFA, 0.1mM beta-NADPH, 0.003% (w/v) BSA.2.Equilibrate to 25℃ and monitor the A340nm until the value is constant using a spectrophotometer.3.Add 2.5ug of recombinant DHFR into reaction mixture and mix immediately. 4.Record the increase in A340nm for 2 minutes |
| Storage: | Can be stored at +4°C short term (1-2 weeks). For long term storage, aliquot and store at -20°C or -70°C. Avoid repeated freezing and thawing cycles. |
| Gene Name: | DHFR dihydrofolate reductase [ Homo sapiens ] |
| Synonyms: | DHFR; dihydrofolate reductase; EC 1.5.1.3 |
| Gene ID: | 1719 |
| mRNA Refseq: | NM_000791 |
| Protein Refseq: | NP_000782 |
| MIM: | 126060 |
| UniProt ID: | P00374 |
| Chromosome Location: | 5q11.2-q13.2 |
| Pathway: | Folate biosynthesis; Metabolic pathways; One carbon pool by folate |
| Function: | NADP or NADPH binding; dihydrofolate reductase activity; oxidoreductase activity |
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