DHFR protein (His tag) (80R-1187)

Purified recombinant Human DHFR protein

Synonyms Dihydrofolate reductase protein, EC 1.5.1.3
Species Human
Protein Type Recombinant
Applications SDS-PAGE
Assay Information 1. Prepare a 3.2 ml reaction mixture in 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 DEg C and monitor at 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 at A340nm for 2 minutes

Images

Coomassie Blue stained SDS-PAGE of DHFR protein (His tag) (80R-1187)

Figure annotation denotes ug of protein loaded and % gel used.

Specifications

Residues 1-187 amino acids: MGSSHHHHHH SSGLVPRGSH MVGSLNCIVA VSQNMGIGKN GDLPWPPLRN EFRYFQRMTT TSSVEGKQNL VIMGKKTWFS IPEKNRPLKG RINLVLSREL KEPPQGAHFL SRSLDDALKL TEQPELANKV DMVWIVGGSS VYKEAMNHPG HLKLFVTRIM QDFESDTFFP EIDLEKYKLL PEYPGVLSDV QEEKGIKYKF EVYEKND
Expression System E.coli
Grade & Purity > 95% pure
Molecular Weight 23.6 kDa (207aa), confirmed by MALDI-TOF.
Tag/Conjugate His tag
Form & Buffer Supplied as a liquid in 20mM Tris-HCl buffer, pH 8.0, containing 0.1M NaCl, 2mM DTT, and 30% glycerol.
Concentration 1 mg/ml

Usage & Assay Information

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

Storage & Safety

Storage Store at 4 deg C for short term storage (1/2 weeks). Aliquot and store at -20 deg C or - 70 deg C for long term storage. Avoid repeated freeze/thaw cycles.

General Information

Biological Significance DHFR, also known as Dihydrofolate reductase, is an enzyme that reduces dihydrofolic acid to tetrahydrofolic acid, using NADPH as electron donor, which can be converted to the kinds of tetrahydrofolate cofactors used in 1-carbon transfer chemistry. Dihydrofolate reductase deficiency has been linked to megaloblastic anemia. Recombinant Dihydrofolate reductase protein, fused to His-tag at N-terminus, was expressed in E. coli and purified by using conventional chromatography techniques.

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Images

  • Coomassie Blue stained SDS-PAGE of DHFR protein (His tag) (80R-1187) | Figure annotation denotes ug of protein loaded and % gel used.

Availability: In stock

Price: £144.23
Size: 100 ug
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