Lactate Dehydrogenase Assay

Shop now

Lactate Dehydrogenase Assay

Method

The reaction velocity is determined by a decrease in absorbance at 340 nm resulting from the oxidation of NADH. One unit causes the oxidation of one micromole of NADH per minute at 25°C and pH 7.3, under the specified conditions.

Reagents

0.2 M Tris⋅HCl, pH 7.3
6.6 mM NADH in above 0.2 M Tris⋅HCl buffer, pH 7.3
30 mM Sodium pyruvate in above 0.2 Tris⋅HCl buffer, pH 7.3

Enzyme

Dissolve at 1 mg/ml in 0.2 M Tris⋅HCl buffer. Dilute enzyme prior to use to obtain a rate of 0.02-0.04 ΔA/min. in Tris buffer and keep cold.

Determination of Protein Concentration:

Procedure

Set spectrophotometer at 340 nm and 25°C.

Pipette into cuvette as follows:

Tris⋅HCl, 0.2 M pH 7.3 2.8 ml

6.6 mM NADH 0.1 ml

30 mM Sodium pyruvate 0.1 ml

Incubate in the spectrophotometer 4-5 minutes to achieve temperature equilibration and establish a blank rate, if any.

Add 0.1 ml of appropriately diluted enzyme and record ΔA₃₄₀/min from initial linear portion.

Calculation

\frac{\text{Units}}{\text{mg}}\text{ = } \frac{\frac{\Delta \text{A}_{340}}{\text{min}}}{\text{6.22 x }\frac{\text{mg enzyme}}{\text{ml reaction mixture}}}

Lactate Dehydrogenase Products

Description

Lactate Dehydrogenase, Lyophilized, Recombinant Rabbit Muscle

Source:

E. coli

Recombinant rabbit muscle Lactate Dehydrogenase produced in E.Coli. Chromatographically purified. A lyophilized powder.

Store at -20°C.

…

Activity

≥250 units per mg protein

Certificate of Analysis

Code

LADCL

Product details

LS002755

5 ku

$124.00

LS002756

25 ku

$575.00

LS002757

Bulk

---

Tris⋅HCl, 0.2 M pH 7.3	2.8 ml
6.6 mM NADH	0.1 ml
30 mM Sodium pyruvate	0.1 ml

Please enter catalog number and quantity

Lactate Dehydrogenase - Assay

Lactate Dehydrogenase Assay

Lactate Dehydrogenase Products