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CS0340

Sigma-Aldrich

Dihydrofolate Reductase Assay Kit

1 kit sufficient for 50-100 tests

Synonym(s):

DHFR Assay Kit

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About This Item

UNSPSC Code:
12161503
NACRES:
NA.84

Quality Level

200

usage

 kit sufficient for 50-100 tests

shipped in

dry ice

storage temp.

−20°C

Gene Information

human ... DHFR(1719)

Application

The kit is used for the detection of DHFR activity and screening of DHFR inhibitors. The assay is based on the ability of DHFR to catalyze the reversible NADPH-dependent reduction of dihydrofolic acid to tetrahydrofolic acid. The reaction progress is followed by monitoring the decrease in absorbance at 340 nm.
Dihydrofolic acid+NADPH+H+ ↔ Tetrahydrofolic acid+NADP+

Biochem/physiol Actions

DHFR (dihydrofolate reductase) is an enzyme that catalyzes a reaction essential for the biosynthesis of nucleotidic bases of DNA. Blocking the enzyme causes cell death as a result of DNA synthesis inhibition. DHFR is an excellent target for anti-tumor drugs.

Features and Benefits

  • Quick and simple method.
  • The kit contains all the reagents required for a colorimetric assay of DHFR activity in cell lysates, tissue homogenates, or column fractions of purified enzyme.
  • The kit includes a purified enzyme for use as a positive control and screening of DHFR inhibitors.
  • The kit includes methotrexate (MTX), a prokaryotic and eukaryotic DHFR specific inhibitor, which exhibits anti-tumor activities.
  • The kit was tested on A431, NIH-3T3, and CHO cell lines, rat liver, kidney, brain, and skeletal muscle tissue extracts, and recombinant DHFR.

Kit Components Only

Product No.
Description
  • Assay Buffer 10x for DHFR 30 mL
  • Dihydrofolate Reductase (DHFR) human .1 U
  • Dihydrofolic acid (DHFR substrate) 3 x 10
  • Amethopterin (+)(methotrexate, MTX)
    (DHFR inhibitor) 2 x 10
  • NADPH (β-Nicotinamide adenine dinucleotide phosphate reduced tetrasodium salt) 25 mg

pictograms

Skull and crossbonesHealth hazard
GHS06,GHS08

signalword

Danger

Hazard Classifications

Acute Tox. 3 Oral - Eye Irrit. 2 - Repr. 1B - Skin Irrit. 2 - STOT SE 3

target_organs

Respiratory system

Storage Class

6.1C - Combustible acute toxic Cat.3 / toxic compounds or compounds which causing chronic effects

flash_point_f

Not applicable

flash_point_c

Not applicable

Certificates of Analysis (COA)

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Amrinder Singh et al.
Scientific reports, 8(1), 3190-3190 (2018-02-18)
We report the first peptide based hDHFR inhibitors designed on the basis of structural analysis of dihydrofolate reductase (DHFR). A set of peptides were rationally designed and synthesized using solid phase peptide synthesis and characterized using nuclear magnetic resonance and
Marta Jorba et al.
Antibiotics (Basel, Switzerland), 10(6) (2021-07-03)
This work reports a detailed characterization of the antimicrobial profile of two trimethoprim-like molecules (compounds 1a and 1b) identified in previous studies. Both molecules displayed remarkable antimicrobial activity, particularly when combined with sulfamethoxazole. In disk diffusion assays on Petri dishes
Bharath Srinivasan et al.
European journal of medicinal chemistry, 103, 600-614 (2015-09-29)
Gram-negative bacteria are implicated in the causation of life-threatening hospital-acquired infections. They acquire rapid resistance to multiple drugs and available antibiotics. Hence, there is the need to discover new antibacterial agents with novel scaffolds. For the first time, this study
James K Martin et al.
Cell, 181(7), 1518-1532 (2020-06-05)
The rise of antibiotic resistance and declining discovery of new antibiotics has created a global health crisis. Of particular concern, no new antibiotic classes have been approved for treating Gram-negative pathogens in decades. Here, we characterize a compound, SCH-79797, that
Prerana Ramadurgum et al.
STAR protocols, 1(2) (2020-10-01)
The use of destabilizing domains (DDs) to conditionally control the abundance of a protein of interest (POI) through a small-molecule stabilizer has gained increasing traction both in vitro and in vivo. Yet there are specific considerations for the development and

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