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CS0340

Dihydrofolate Reductase Assay Kit

1 kit sufficient for 50-100 tests

Synonyme(s) :

DHFR Assay Kit

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1 kit
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720,00 $

A propos de cet article

NACRES:
NA.84
UNSPSC Code:
12161503

720,00 $

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Quality Segment

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.

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Cet article
D6566WH0001719M1D1067
Dihydrofolate Reductase Assay Kit 1 kit sufficient for 50-100 tests

Sigma-Aldrich

CS0340

Dihydrofolate Reductase Assay Kit

Dihydrofolate Reductase human ≥80% (SDS-PAGE), recombinant, expressed in E. coli, ≥1 units/mg protein

Sigma-Aldrich

D6566

Dihydrofolate Reductase human

Monoclonal Anti-DHFR antibody produced in mouse clone 2B10, purified immunoglobulin, buffered aqueous solution

Sigma-Aldrich

WH0001719M1

Monoclonal Anti-DHFR antibody produced in mouse

Anti-DHFR, N-terminal antibody produced in rabbit ~1.0 mg/mL, affinity isolated antibody, buffered aqueous solution

Sigma-Aldrich

D1067

Anti-DHFR, N-terminal antibody produced in rabbit

description

1 kit sufficient for 50-100 tests

description

≥80% (SDS-PAGE), recombinant, expressed in E. coli, ≥1 units/mg protein

description

clone 2B10, purified immunoglobulin, buffered aqueous solution

description

~1.0 mg/mL, affinity isolated antibody, buffered aqueous solution

Gene Information

human ... DHFR(1719)

Gene Information

human ... DHFR(1719)

Gene Information

human ... DHFR(1719)

Gene Information

human ... DHFR(1719)
mouse ... Dhfr(13361)
rat ... Dhfr(24312)

Quality Level

200

Quality Level

200

Quality Level

100

Quality Level

200

usage

 kit sufficient for 50-100 tests

usage

-

usage

-

usage

-

shipped in

dry ice

shipped in

wet ice

shipped in

dry ice

shipped in

dry ice

storage temp.

−20°C

storage temp.

−20°C

storage temp.

−20°C

storage temp.

−20°C

Composants de kit seuls

Réf. du produit
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

Pictogrammes

Skull and crossbonesHealth hazard
GHS06,GHS08

Mot-clé

Danger

Classe de stockage

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

Point d'éclair (°F)

Not applicable

Point d'éclair (°C)

Not applicable

Que se passe-t-il ?

WGK 3

Hazard Classifications

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

Organes cibles

Eyes,Central nervous system, Respiratory system

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Certificats d'analyse (COA)

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Questions

1–2 of 2 Questions  

  1. 1. what is the origin of this DHFR. Is it from Human, E. coli or any other. 2. I want to relate antibacterial activity of my compound with this enzyme assay kit. Is it obvious.

    1 answer

    1. The Dihydrofolate Reductase that is provided in the kit is product D6566, a recombinant human enzyme that is expressed in E. coli. Please see the link below to review the datasheet for this component:
      https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/213/424/d6566dat.pdf

      One intended use of this kit is for screening DHFR inhibitors, including inhibitors of bacterial DHFR. This kit is expected to be suitable for the measurement of antibacterial activity. This characteristic is expressed as DHFR inhibitory activity of a compound of interest. Please see the link below to review the datasheet for this kit:
      https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/427/408/cs0340bul.pdf

      Helpful?

  2. I want to measure activity in protein variants that have different cellular concentration (protein stability), will this affect my activity?

    1 answer

    1. It is unclear the relation between cellular concentration and protein stability. It seems the intention is to produce dihydrofolate reductase recombinantly and then measure the activity. Protein stability is dependent on several different factors such as pH, temperature, buffering conditions, storage, purification, etc. If a protein is not stable it can certainly affect the activity. Testing would be necessary to determine the overall impact on the activity as you optimize your process.

      To further discuss this application, we kindly ask you to navigate to the link https://www.sigmaaldrich.com/techservice, and click on "Product Technical Inquires" under the Products Section with all the required information so that a member of our team can reach out to you to assist further. Thank you.

      Helpful?

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