Malate Dehydrogenase (MDH), CAS No.9001-64-3

CAS: 9001-64-3 Cat. No.: R1506880 Peso molecular: 70 kD Número EC: 232-622-5

GRADE & PURITY Bioactive, Recombinant, ActiBioPure™, High Performance, EnzymoPure™, ≥40 U/mg enzyme powder; expressed in E.coli

Accession #

P61889

Expression system

E. coli

Bioactivity

≥40 U/mg enzyme powder

Descripción general

  Malate dehydrogenase (MDH) is a key oxidoreductase, which catalyzes the reversible conversion of malate and oxaloacetic acid in cell metabolism and participates in the TCA cycle and energy metabolism.

Application: This enzyme is useful for enzymatic determination of L-malate and of glutamate oxaloacetate transaminase (GOT) in clinical analysis.

Assay Procedure

Principle:

Reagents:

A.K-Phosphate buffer: 0.1M, pH7.5;

B.Oxaloacetate solution: 15mM [2.0mg oxaloacetic acid (MW=132.1)/ml of ice-cold K-phosphate buffer(A), This reagent is rather unstable and should be store in an ice-bath during use](Should be prepared fresh)

C.NADH solution: 6.0mM [4.25mg NADH·2Na,(MW=709.4 )/ml of H₂O] (Should be prepared fresh)

D.Enzyme diluent: 0.1M K-Phosphate buffer,pH7.5 contg.0.2% BSA

Procedure:

1.Prepare the following reaction mixture in a cuvette (d=1.0 cm) and equilibrate at 30℃ for about 5 minutes. 

2.8ml	K-Phosphate buffer , pH7.5	( A )
0.1ml	Oxaloacetate solution	( B )
0.1ml	NADH solution	( C )

Concentration in assay mixture

K-Phosphate buffer	97mM
Oxaloacetate	0.49mM
NADH	0.20mM

2.Add 0.05ml of the enzyme solution* and mix by gentle inversion.

3.Record the decrease in optical density at 340nm against water for 3~4minutes in a spectrophotometer thermostated at 30℃,and calculate the △OD per minute from the initial linear portion of the curve ( △OD test).

At the same time,measure the blank rate (△OD Blank) by using the same method as the test expect that the enzyme diluent is added instead of the enzyme solution.

*:Dissolve the enzyme preparation in ice-cold enzyme diluent(D)and dilute to 0.05~0.5U/ml with the same buffer and store on ice.

Calcuation:

Activity can be calculated by using the following formula:

Vt: Total volume(3.05ml)

Vs: Sample volume(0.05ml)

6.22: Millimolar extinction coefficient of NADH under the assay condition (cm²/μM)

1.0: Light path length(cm)

df: Dilution factor

C: Enzyme concentration in dissolution(c mg/ml)

Specifications

Product Name

Malate Dehydrogenase (MDH), CAS No.9001-64-3

Sinónimos

(S)-malate dehydrogenase (NAD(+), oxaloacetate-forming) | malate dehydrogenase | malic dehydrogenase | (S)-malate:NAD+ oxidoreductase | (S)-malate dehydrogenase (NAD+, oxaloacetate-forming) | malic dehydrogenase (ambiguous) | L-malate dehydrogenase (ambig

Grado

ActiBioPure™, Bioactive, High Performance, Recombinant, EnzymoPure™

Especificaciones y pureza

Bioactive, ActiBioPure™, High Performance, EnzymoPure™, Recombinant, ≥40 U/mg enzyme powder; expressed in E.coli

Bioactividad

≥40 U/mg enzyme powder

Secuencia N-terminal

One unit causes the oxidation of one micromole of NADH per minute at pH7.5 at 30°C.

Número de adhesión

P61889

CAS

9001-64-3

Número de la Comisión de Enzimas

EC 1.1.1.37

Tipo de molécula

Enzyme

Almacenamiento y envío

Concentración

≥40 U/mg enzyme powder; expressed in E.coli

Reconstitución

Reconstitute in pure water.

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℃ long term (12 months). Upon reconstitution, it is recommended to aliquot. Avoid freeze/thaw cycle.