CS0340

Sigma-Aldrich

Dihydrofolate Reductase Assay Kit

 kit sufficient for 50-100 tests

Synonym(s):
DHFR Assay Kit
NACRES:
NA.84
Pricing and availability is not currently available.

Quality Level

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

Signal Word

Danger

Target Organs

Respiratory system

Hazard Codes

T

Risk Statement

46-61-25-36/37/38

Safety Statement

53-26-45

RIDADR

UN 3316 9

WGK Germany

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Certificate of Analysis
Certificate of Origin
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.
Cell chemical biology, 27(5), 622-634 (2020-04-25)
Destabilizing domains (DDs), such as a mutated form of Escherichia coli dihydrofolate reductase (ecDHFR), confer instability and promote protein degradation. However, when combined with small-molecule stabilizers (e.g., the antibiotic trimethoprim), DDs allow positive regulation of fusion protein abundance. Using a combinatorial screening...
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