DHFR Mouse

Dihydrofolate Reductase Mouse Recombinant
Cat. No.
BT18461
Source
Escherichia Coli.
Synonyms
Dihydrofolate reductase, DHFR, DHFRP1, AA607882, AI662710, AW555094, 8430436I03Rik.
Appearance
Sterile Filtered colorless solution.
Purity
Greater than 95.0% as determined by SDS-PAGE.
Usage
THE BioTek's products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.
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Description

DHFR Mouse Recombinant fused with a 20 amino acid His tag at N-terminus produced in E.Coli is a single, non-glycosylated, polypeptide chain containing 207 amino acids (1-187 a.a.) and having a molecular mass of 23.8kDa. The DHFR is purified by proprietary chromatographic techniques.

Product Specs

Introduction
Dihydrofolate reductase (DHFR) is an enzyme that catalyzes the reduction of dihydrofolic acid to tetrahydrofolic acid. This reaction is crucial as it utilizes NADPH as an electron donor to produce tetrahydrofolate cofactors, essential components in one-carbon transfer chemistry. Tetrahydrofolate is vital for the de novo synthesis of purines, thymidylic acid, and certain amino acids. While the functional DHFR gene is located on chromosome 5, multiple processed pseudogenes and dihydrofolate reductase-like genes are found on other chromosomes. A deficiency in DHFR can lead to megaloblastic anemia. Furthermore, studies show that DHFR knockdown contributes to the anticancer properties of 2-hydroxyoleic acid. Additionally, variations in serum and red blood cell folate levels in women have been linked to polymorphisms in the DHFR gene, including insertions and deletions.
Description
This product is a recombinant mouse Dihydrofolate reductase (DHFR) protein with a 20 amino acid His tag attached to its N-terminus. It is expressed in E. coli and has a molecular weight of 23.8 kDa. The protein is a single, non-glycosylated polypeptide chain consisting of 207 amino acids (with amino acids 1-187 representing the DHFR sequence). Purification is achieved using proprietary chromatographic techniques.
Physical Appearance
The product is a sterile, colorless solution.
Formulation
The DHFR solution is provided at a concentration of 1 mg/ml and contains the following components: 20mM Tris-HCl buffer (pH 8.0), 10% glycerol, 2mM DTT, and 0.1M NaCl.
Stability
For short-term storage (up to 4 weeks), the product can be stored at 4°C. For long-term storage, it is recommended to freeze the product at -20°C. To ensure optimal stability during long-term storage, consider adding a carrier protein such as HSA or BSA (0.1%). It's essential to avoid repeated freezing and thawing of the product.
Purity
The purity of this product is greater than 95% as determined by SDS-PAGE analysis.
Biological Activity
The specific activity of this product is greater than 0.2 units/mg. One unit is defined as the amount of enzyme that catalyzes the conversion of 1.0 µmol of 7,8-dihydrofolate and beta-NADPH to 5,6,7,8-tetrahydrofolate and beta-NADP per minute at a pH of 6.5 and a temperature of 25°C.
Synonyms
Dihydrofolate reductase, DHFR, DHFRP1, AA607882, AI662710, AW555094, 8430436I03Rik.
Source
Escherichia Coli.
Amino Acid Sequence
MGSSHHHHHH SSGLVPRGSH MVRPLNCIVA VSQNMGIGKN GDLPWPPLRN EFKYFQRMTT TSSVEGKQNL VIMGRKTWFS IPEKNRPLKD RINIVLSREL KEPPRGAHFL AKSLDDALRL IEQPELASKV DMVWIVGGSS VYQEAMNQPG HLRLFVTRIM QEFESDTFFP EIDLGKYKLL PEYPGVLSEV QEEKGIKYKF EVYEKKD.

Product Science Overview

Structure and Function

DHFR is a small enzyme, approximately 21 kDa in size . It is highly conserved across different species, including humans and mice. The enzyme’s primary function is to maintain the cellular levels of tetrahydrofolate, a cofactor required for one-carbon transfer reactions . These reactions are crucial for the synthesis of nucleotides and amino acids, making DHFR indispensable for cell growth and division .

Recombinant DHFR

Recombinant DHFR refers to the enzyme produced through recombinant DNA technology. This involves inserting the DHFR gene into a suitable expression system, such as E. coli, to produce the enzyme in large quantities . Recombinant DHFR is often tagged with a histidine tag to facilitate purification and is used in various biochemical assays and research applications .

Applications in Research

Recombinant DHFR is widely used in research to study its role in cellular metabolism and its potential as a target for chemotherapy drugs. Inhibitors of DHFR, such as methotrexate, are used to treat certain types of cancer and autoimmune diseases by blocking the enzyme’s activity, thereby inhibiting DNA synthesis and cell proliferation .

Importance in Drug Development

The study of DHFR, particularly its recombinant forms, is crucial in drug development. By understanding how DHFR functions and how it can be inhibited, researchers can develop more effective drugs with fewer side effects. The enzyme’s role in folate metabolism also makes it a target for antibiotics and antimalarial drugs .

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