DHFR / DHFRP1 (1-187, His-tag) Human Protein

Specifications

Product Data
Species	Human
Expression Host	E. coli
Expression cDNA Clone or AA Sequence	Protein Sequence (showhide) MGSSHHHHHH SSGLVPRGSH MVGSLNCIVA VSQNMGIGKN GDLPWPPLRN EFRYFQRMTT TSSVEGKQNL VIMGKKTWFS IPEKNRPLKG RINLVLSREL KEPPQGAHFL SRSLDDALKL TEQPELANKV DMVWIVGGSS VYKEAMNHPG HLKLFVTRIM QDFESDTFFP EIDLEKYKLL PEYPGVLSDV QEEKGIKYKF EVYEKND
Tag	His-tag
Predicted MW	23.6 kDa
Concentration	lot specific
Purity	>95% by SDS - PAGE
Buffer	Presentation State: Purified State: Liquid purified protein Buffer System: 20 mM Tris-HCl buffer (pH 8.0) containing 0.1 M Nacl, 2 mM DTT, and 30% glycerol
Preparation	Liquid purified protein
Protein Description	Recombinant Dihydrofolate reductase protein, fused to His-tag at N-terminus, was expressed in E.coli and purified by using conventional chromatography techniques.
Storage	Store undiluted at 2-8°C for up to two weeks or (in aliquots) at -20°C or -70°C for longer. Avoid repeated freezing and thawing.
Stability	Shelf life: one year from despatch.
Bioactivity	Biological: 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. 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°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)
Reference Data
RefSeq	NP_000782
Locus ID	1719
UniProt ID	P00374, B0YJ76
Cytogenetics	5q14.1
Synonyms	DHFRP1; DYR
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

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