Carboxypeptidase B Assay

Shop now

Carboxypeptidase B Assay

Method

Activity is measured by the spectrophotometric method of Folk et al. (1960) where the reaction velocity is determined by an increase in absorbance at 254 nm resulting from the hydrolysis of hippuryl-L-arginine. One unit causes the hydrolysis of one micromole of hippuryl-L-arginine per minute at 25°C and pH 7.65 under the specified conditions.

Reagents

0.025 M Tris⋅HCl buffer, pH 7.65 containing 0.1 M sodium chloride
0.001 M Hippuryl-L-arginine in 0.025 M Tris⋅HCl pH 7.65 containing 0.1 M sodium chloride

Enzyme

Dilute stock solution with reagent grade water to a concentration of 1-5 units/ml.

Procedure

Set spectrophotometer at 254 nm and 25°C.

Pipette 2.9 ml of substrate into cuvette and incubate in spectrophotometer at 25°C for 3-4 minutes to reach temperature equilibration and establish blank rate, if any. Add 0.1 ml of diluted enzyme and record increase in A₂₅₄ for 3-4 minutes. Determine ΔA₂₅₄/minute from the initial linear portion of the curve.

Calculation

where 0.349 is the extinction coefficient of hippuric acid formed during the reaction

\frac{\text{Units}}{\text{mg}} = \frac{\frac{\Delta \text{A}_{254}}{\text{min}}}{\frac{\text{0.349 x mg enzyme}}{\text{ml reaction mixture}}}

Carboxypeptidase B Products

Description

Carboxypeptidase B

Source:

Porcine Pancreas

Chromatographically purified. A solution in 100mM sodium chloride. Chymotrypsin and trypsin 0.02%.

Store at -20°C.

…

Ice Pack required

Activity

≥170 units per mg protein

Certificate of Analysis

Code

COBC

Product details

LS005305

5 mg

$96.00

LS005301

10 mg

$167.00

LS005304

50 mg

$710.00

LS005302

Bulk

---

Please enter catalog number and quantity

Carboxypeptidase B - Assay

Carboxypeptidase B Assay

Carboxypeptidase B Products