CS0340

Dihydrofolate Reductase Assay Kit

1 kit sufficient for 50-100 tests

• Synonym(s): DHFR Assay Kit
• UNSPSC Code: 12161503
• NACRES: NA.84

Properties

• Quality Level: 200
• usage: kit sufficient for 50-100 tests
• shipped in: dry ice
• storage temp.: −20°C
• Gene Information: human ... DHFR(1719)

Description

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

• 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

Safety Information

• Pictograms: GHS06,GHS08
• Signal Word: Danger
• Hazard Statements: H301,H315,H319,H335,H360FD
• Precautionary Statements: P202 - P261 - P264 - P301 + P310 - P302 + P352 - P305 + P351 + P338
• Hazard Classifications: Acute Tox. 3 Oral - Eye Irrit. 2 - Repr. 1B - Skin Irrit. 2 - STOT SE 3
• Target Organs: Respiratory system
• Storage Class Code: 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