Recombinant Human DHFR Protein

Cat. No.: rp188305

GRADE & PURITY Carrier Free, Bioactive, ActiBioPure™, His Tag, ≥95%(SDS-PAGE), See COA

Accession #

P00374

Protein Tag

C-His

Expression system

E. coli

Endotoxin Concentration

<1.0 EU/μg

Bioactivity

1. Measured by the reduction of dihydrofolic acid (DHF). The specific activity is >5,500 pmol/min/μg, as measured under the described conditions. 2. Immobilized Recombinant Human DHFR Protein (rp188305) at 1.0 μg/mL can bind Recombinant Dihydrofolate reductase/DHFR Antibody (Ab099867) with the EC50 of 4.9 ng/mL.

Specifications

Product Name

Recombinant Human DHFR Protein

Sinónimos

DHFR | DHFRP1 | Dihydrofolate Reductase | DYR | EC 1.5.1.3

Grado

ActiBioPure™, Bioactive, Carrier Free, His Tag

Especificaciones y pureza

Carrier Free, Bioactive, ActiBioPure™, His-Tag, ≥95%(SDS-PAGE), See COA

Mecanismos bioquímicos y fisiológicos

Dihydrofolate Reductase (DHFR) is an approximately 21 kDa enzyme that catalyzes the NADPH-dependent reduction of dihydrofolate to tetrahydrofolate, which is crucial for the synthesis of purines, thymidylate, and certain amino acids. Structurally, DHFR con

Bioactividad

1. Measured by the reduction of dihydrofolic acid (DHF). The specific activity is >5, 500 pmol/min/μg, as measured under the described conditions. 2. Immobilized Recombinant Human DHFR Protein (rp188305) at 1.0 μg/mL can bind Recombinant Dihydrofolate reductase/DHFR Antibody (Ab099867) with the EC50 of 4.9 ng/mL.

Concentración de endotoxinas

<1.0 EU/μg

Aminoácidos

1-187 aa

Secuencia

MVGSLNCIVAVSQNMGIGKNGDLPWPPLRNEFRYFQRMTTTSSVEGKQNLVIMGKKTWFSIPEKNRPLKGRINLVLSRELKEPPQGAHFLSRSLDDALKLTEQPELANKVDMVWIVGGSSVYKEAMNHPGHLKLFVTRIMQDFESDTFFPEIDLEKYKLLPEYPGVLSDVQEEKGIKYKFEVYEKNDHHHHHH

Número de adhesión

P00374

Peso molecular previsto

22.2 kDa

SDS-PAGE

23.5 kDa, under reducing conditions; 23.5 kDa, under non-reducing conditions.

Tipo de molécula

Protein

Almacenamiento y envío

Concentración

See COA

Condiciones de almacenamiento de almacenamiento

Store at -20°C,Avoid repeated freezing and thawing

Enviado en

Ice chest + Ice pads

Estabilidad y almacenamiento

Store at -20°C for 1 year. Avoid freeze/thaw cycle.

Imágenes

ELISA: Recombinant Human DHFR Protein (rp188305)

Recombinant Human DHFR Protein (rp188305) - ELISA
Immobilized Recombinant Human DHFR Protein (rp188305) at 1.0 μg/mL can bind Recombinant Dihydrofolate reductase/DHFR Antibody (Ab099867) with the EC50 of 4.9 ng/mL.

SDS-PAGE: Recombinant Human DHFR Protein (rp188305)

Recombinant Human DHFR Protein (rp188305) - SDS-PAGE
2 μg/lane of Recombinant Human DHFR Protein was resolved with SDS-PAGE under reducing (R) and non-reducing (N) conditions and visualized by Coomassie® Blue staining, showing a band at 23.5 kDa.

Enzyme Activity: Recombinant Human DHFR Protein (rp188305Z)

Recombinant Human DHFR Protein - Enzyme Activity
Measured by the reduction of dihydrofolic acid (DHF). The specific activity is >5,500 pmol/min/μg, as measured under the described conditions.
Materials:
Assay Buffer: 50 mM MES, 25 mM Tris, 100 mM NaCl, 25 mM Ethanolamine, 2 mM DTT
Recombinant Human DHFR Protein (rp188305Z)
beta -Nicotinamide adenine dinucleotide phosphate reduced, tetrasodium salt ( beta -NADPH) (N276326), 10 mM stock in deionized water
Dihydrofolic acid (DHF)  (D135507), 10 mM stock in Assay Buffer + 4.5 mM NaOH
Procedures：
1.Dilute rhDHFR to 0.33 μg/mL in Assay Buffer.
2. Prepare a Substrate Mixture containing 0.8 mM DHF and 1 mM beta -NADPH in Assay Buffer.
3. Load 50 μL of 0.33 μg/mL rhDHFR into a plate, and start the reaction by adding 50 μL of Substrate Mixture.  Include a Substrate Blank containing 50 μL of Assay Buffer and 50 μL of Substrate Mixture.
4. Read at an absorbance of 339 nm in kinetic mode for 5 minutes.
5. Calculate specific activity:

*Adjusted for Substrate Blank 

**Using the extinction coefficient 6220 M⁻¹cm⁻¹ 

***Using the path correction 0.32 cmNote: the output of many spectrophotometers is in mOD.
Per Well:
rhDHFR: 0.0165 μg, DHF: 0.4 mM, beta -NADPH: 0.5 mM