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AR09254PU-L DHFR / DHFRP1 (1-187, His-tag)

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0.5 mg / €600.00
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Quick Overview

DHFR / DHFRP1

AR09254PU-L

Product Description for DHFR / DHFRP1

DHFR / DHFRP1.
Presentation: Purified

Properties for DHFR / DHFRP1

Product Category Proteins & Growth Factors
Quantity 0.5 mg
Synonyms Dihydrofolate reductase
Presentation Purified
Molecular weight 23.6 kDa
Source E. coli
Species (Protein) Human
Shipping to Worldwide
PDF datasheet View Datasheet
Manufacturer OriGene Technologies GmbH
Material safety datasheet MSDS for Proteins (de)

Datasheet Extract

Immunogen
Swiss Prot Num:
P00374
Purity > 95 % by SDS - PAGE
Background 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.
Concentration 1.0 mg/ml (determined by Bradford assay)
General Readings
  1. Loveridge EJ, Rodriguez RJ, Swanwick RS, Allemann RK. Effect of dimerization on the stability and catalytic activity of dihydrofolate reductase from the hyperthermophile Thermotoga maritima. Biochemistry. 2009 Jun 30;48(25):5922-33. doi: 10.1021/bi900411a. PubMed PMID: 19453185.
  2. Hu Y, Kireev I, Plutz M, Ashourian N, Belmont AS. Large-scale chromatin structure of inducible genes: transcription on a condensed, linear template. J Cell Biol. 2009 Apr 6;185(1):87-100. doi: 10.1083/jcb.200809196. PubMed PMID: 19349581. (Free PMC Article available, 7 images available)
  3. Hillcoat BL, Nixon PF, Blakley RL. Effect of substrate decomposition on the spectrophotometric assay of dihydrofolate reductase. Anal Biochem. 1967 Nov;21(2):178-89. PubMed PMID: 4384177.
  4. Peterson DL, Gleisner JM, Blakley RL. Bovine liver dihydrofolate reductase: purification and properties of the enzyme. Biochemistry. 1975 Dec 2;14(24):5261-7. PubMed PMID: 45.
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.
Shelf life: one year from despatch.
Description
Description:
Recombinant Dihydrofolate reductase protein, fused to His-tag at N-terminus, was expressed in E.coli and purified by using conventional chromatography techniques.
AA Sequence:
MGSSHHHHHH SSGLVPRGSH MVGSLNCIVA VSQNMGIGKN GDLPWPPLRN EFRYFQRMTT TSSVEGKQNL VIMGKKTWFS IPEKNRPLKG RINLVLSREL KEPPQGAHFL SRSLDDALKL TEQPELANKV DMVWIVGGSS VYKEAMNHPG HLKLFVTRIM QDFESDTFFP EIDLEKYKLL PEYPGVLSDV QEEKGIKYKF EVYEKND
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°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.
Molecular weight:
(207 aa), confirmed by MALDI-TOF
Format
Buffer System:
20 mM Tris-HCl buffer (pH 8.0) containing 0.1 M Nacl, 2 mM DTT, and 30% glycerol
Endotoxin level:
< 1.0 EU per 1 μg of protein (determined by LAL method)
Purity:
>95% by SDS - PAGE
State:
Liquid purified protein
Purified
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