Recombinant Human Dihydrofolate Reductase, His-tagged

• 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).

Specification

• Species: Human
• Source: E.coli
• Tag: His
• 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.
• 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.
• Publications:
  Human dihydrofolate reductase influences the sensitivity of the malaria parasite Plasmodium falciparum to ketotifen–A cautionary tale in screening transgenic parasites (2016)

Gene Information

• 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