DHFR / DHFRP1 (1-187, His-tag) Human Protein
Recombinant Dihydrofolate reductase protein, fused to His-tag at N-terminus, was expressed in E.coli and purified by using conventional chromatography techniques.
|Description||Recombinant Dihydrofolate reductase protein, fused to His-tag at N-terminus, was expressed in E.coli and purified by using conventional chromatography techniques.|
|Expression Host||E. coli|
|Expression cDNA Clone or AA Sequence||MGSSHHHHHH SSGLVPRGSH MVGSLNCIVA VSQNMGIGKN GDLPWPPLRN EFRYFQRMTT TSSVEGKQNL VIMGKKTWFS IPEKNRPLKG RINLVLSREL KEPPQGAHFL SRSLDDALKL TEQPELANKV DMVWIVGGSS VYKEAMNHPG HLKLFVTRIM QDFESDTFFP EIDLEKYKLL PEYPGVLSDV QEEKGIKYKF EVYEKND|
|Predicted MW||23.6 kDa|
|Concentration||1.0 mg/ml (determined by Bradford assay)|
|Purity||> 95 % by SDS - PAGE|
|Buffer||20 mM Tris-HCl buffer (pH 8.0) containing 0.1 M Nacl, 2 mM DTT, and 30% glycerol|
|Bioactivity||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°C.
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°C and monitor the A340nm until the value is constant using a spectrophotometer.
3. Add 2.5 ug of recombinant DHFR into reaction mixture and mix immediately.
4. Record the increase in A340nm for 2 minutes.
|Endotoxin||< 1.0 EU per 1 µg of protein (determined by LAL method)|
|Preparation||Liquid purified protein|
|Summary||Dihydrofolate reductase converts dihydrofolate into tetrahydrofolate, a methyl group shuttle required for the de novo synthesis of purines, thymidylic acid, and certain amino acids. While the functional dihydrofolate reductase gene has been mapped to chromosome 5, multiple intronless processed pseudogenes or dihydrofolate reductase-like genes have been identified on separate chromosomes. Dihydrofolate reductase deficiency has been linked to megaloblastic anemia. Several transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq, Mar 2014]|
|Protein Families||Druggable Genome, Stem cell - Pluripotency|
|Protein Pathways||Folate biosynthesis, Metabolic pathways, One carbon pool by folate|