Dihydrofolate Reductase Assay Kit
SIGMA/CS0340 - 1 kit sufficient for 50-100 tests
Synonym: DHFR Assay Kit
Product Type: Chemical
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Unit Definition One unit will convert 1.0 μmole of dihydrofolic acid to tetrahydrofolic acid in 1 minute at pH 7.5 at 22 °C. Stoichiometry: Tetrahydrofolate + NADP+ → Dihydrofolate + NADPH + H+
| Quality Level | 200  |
| shipped in | dry ice |
| storage temp. | −20°C |
| usage | kit sufficient for 50-100 tests |
| 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. |
| Symbol |   GHS06,GHS08 |
| Signal word | Danger |
| Hazard statements | H301 - H315 - H319 - H335 - H360 |
| Precautionary statements | P201 - P301 + P310 + P330 - P302 + P352 - P305 + P351 + P338 |
| Hazard Codes | T |
| Risk Statements | 46-61-25-36/37/38 |
| Safety Statements | 53-26-45 |
| RIDADR | UN 3316 9 |
| WGK Germany | WGK 3 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | −20°C |
| UNSPSC | 12161503 |
| Components | Assay Buffer 10x for DHFR 30 mL; Dihydrofolate Reductase (DHFR) human .1 U; Dihydrofolic acid (DHFR substrate) 3 x 10 mg; Amethopterin (+)(methotrexate, MTX) (DHFR inhibitor) 2 x 10 mg; NADPH (β-Nicotinamide adenine dinucleotide phosphate reduced tetrasodium salt) 25 mg |