EC E N Z Y Cholesterol Oxidase M ES (Recombinant) · MES BICINE MOPS CHES KPO4 Na ACETATE PHOSPHATE...

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ORIGIN Microbial SPECIFICATIONS Appearance Yellow free flowing powder Activity >32 U/mg powder at 37°C Specific Activity >45 U/mg protein at 37°C Contaminants Catalase <1% Glucose Oxidase <0.01% Uricase <0.01% UNIT DEFINITION One unit of activity is defined as the amount of enzyme that will catalyse the oxidation of 1.0 micromole of cholesterol per minute at 37°C under the standard assay method conditions. Refer to Table 1 for guidance on factors to adjust units according to temperature of assay. Cholesterol Oxidase (Recombinant) ENZYMES CAT# RECO-70-1229 EC# 1.1.3.6 CAT# RECO-70-1229 EC# 1.1.3.6 ENZYMES ASSAY TEMPERATURE 25°C 0.75 30°C 0.86 37°C 1.00 45°C 1.02 50°C 1.01 FACTOR RELATIVE TO 37° RESULT TABLE 1: TEMPERATURE FACTORS FOR UNIT CONVERSION Note: Temperature can influence the level of available oxygen in the reaction mixture. ASSAY PRINCIPLE Cholesterol Oxidase (CO) catalyses the following reaction: Cholesterol + O 2 Cholestenone + H 2 O 2 CO

Transcript of EC E N Z Y Cholesterol Oxidase M ES (Recombinant) · MES BICINE MOPS CHES KPO4 Na ACETATE PHOSPHATE...

Page 1: EC E N Z Y Cholesterol Oxidase M ES (Recombinant) · MES BICINE MOPS CHES KPO4 Na ACETATE PHOSPHATE AMPSO MES TRIS CHES MOPS BICINE CAPS. Title: Layout 1 Created Date: 20170324094319-08'00'

O R I G I N Microbial

� S P E C I F I C A T I O N SAppearance Yellow free flowing powderActivity >32 U/mg powder at 37°CSpecific Activity >45 U/mg protein at 37°CContaminants Catalase <1%

Glucose Oxidase <0.01%Uricase <0.01%

� U N I T D E F I N I T I O NOne unit of activity is defined as the amount of enzyme that will catalyse the oxidation of 1.0micromole of cholesterol per minute at 37°C under the standard assay method conditions.Refer to Table 1 for guidance on factors to adjust units according to temperature of assay.

Cholesterol Oxidase(Recombinant)

E N Z Y M E S

CAT# RECO-70-1229

EC# 1.1.3.6

C A T # R E C O - 7 0 - 1 2 2 9E C # 1 . 1 . 3 . 6

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ASSAY TEMPERATURE

25°C 0.75

30°C 0.86

37°C 1.00

45°C 1.02

50°C 1.01

FACTOR RELATIVE TO 37° RESULT

TABLE 1 : T EMPERATURE FACTORS FOR UN I T CONVERS ION

Note: Temperature can influence the level of available oxygen in the reaction mixture.

� A S S AY P R I N C I P L ECholesterol Oxidase (CO) catalyses the following reaction:

Cholesterol + O2 Cholestenone + H2O2CO

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CAT# RECO-70-1229

EC# 1.1.3.6

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SUBS TRATE SPEC I F IC I TY

SUBSTRATE

Pregnenolone 104%

Cholesterol 100%

β-Cholestanol 81%

Stigmasterol 51%

Ergosterol 39%

% OF CHOLESTEROL ACTIVITY

SUBSTRATE

Cholesteryl Linoleate 0.02%

Cholecalciferol 0.001%

Androsterone 0.001%

Estradiol 0.0003%

% OF CHOLESTEROL ACTIVITY

Molecular Weight (by SDS Page): 58kD

Kmvalue (using Eadie-Hofstee): 5 x 10-6 M (Cholesterol)

Optimum pH (Fig. 1): pH 6.5 to 7.5

Optimum Temperature (Fig. 2): 45°C

Stable pH Range (Fig. 3): pH 4.5 to 9.0 (25°C for 20 hrs)

Thermal Stability (Fig. 4): Stable at 60°C and below (pH 7.0 for 15 mins)

Substrate specificity was tested in-house by replacing cholesterol with alternativesubstrates in the CO assay i.e. at 0.1mM concentration.

� C H A R A C T E R I S T I C SThis recombinant grade of CO is highly purified. Its main characteristics are as follows:

� A P P L I C A T I O NCholesterol Oxidase can be used in combination with Cholesterol Esterase (inbiosensors, test strips or other clinical chemistry formats) to determine the level ofcholesterol in blood. It is highly soluble over a wide pH.

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F IGURE 3 : pH S TAB I L I TY

CAT# RECO-70-1229

EC# 1.1.3.6

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[email protected]

THE AMERICASSekisui Diagnostics, LLC4 Hartwell Place Lexington, MA 02421 Phone: 800 332 1042Fax: 800 762 6311

INTERNATIONALSekisui Diagnostics (UK) LimitedLiphook Way, AllingtonMaidstone, Kent, ME16 0LQ, UKPhone: +44 1622 607800Fax: +44 1622 607801

AP-00-0019.1.1 03/17© 2017 Sekisui Diagnostics, LLC. All rights reserved. Because every result matters™ is a trademark of Sekisui Diagnostics, LLC.

RELATIVE ACTIVITY (%)

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F IGURE 1 : OP T IMUM pH

F IGURE 4 : T EMPERATURE S TAB I L I TY

F IGURE 2 : OP T IMUM T EMPERATURE

pH

3 4 5 6 7 8 9 10 11

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TEMPERATURE (°C)

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-20

RELATIVE ACTIVITY (%)

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