K-ACETAF，乙酸[AF法]检测试剂盒，ACETIC ACID(Acetyl-CoA Synthetase Format)assay Kit

K-ACETAF，乙酸[AF法]检测试剂盒，ACETIC ACID(Acetyl-CoA Synthetase Format)assay Kit

产品识别号：K-ACETAF

产品品名：乙酸[AF法]检测试剂盒

英文品名：ACETIC ACID(Acetyl-CoA Synthetase Format)assay Kit

规格型号：

(141.6 mL of reagent [R1 + R2] per kit;

equivalent to 456 reactions of 0.31 mL)

INTRODUCTION:

The most widely used method for enzymatic quantification of acetic acidis that based on the use of acetyl-coenzyme A synthetase (ACS),according to equations 1 – 3 below. For auto-analyser applications itbecomes necessary to prepare a “master mix” reagent (R1), containing allcomponents of the assay except the reaction initiation enzyme, ACS.However, reagent R1 when prepared from some kits has very limitedon-machine stability, owing to a rapidly increasing absorbance value. Toovercome this issue, Megazyme developed this kit (K-ACETAF), thatdoesn’t exhibit this instability phenomenon. Additionally, theanti-inhibitory compound polyvinylpyrollidone (PVP) has also beenincorporated into the assay to prevent inhibition caused by certaintannins found in grape juice and wine.

KITS:

Kits suitable for the preparation of 141.6 mL of reagent (equivalent to456 reactions of 0.31 mL) are available from Megazyme.

The kits contain the full assay method plus:

Bottle 1: Buffer (30 mL, pH 8.4) plus L-malic acid, PVP and sodiumazide (0.02 % w/v) as a preservative. Stable for > 2 years at 4°C.

Bottle 2: (x2) NAD+ plus ATP and CoA.Freeze dried powder.Stable for> 5 years at -20°C.

Bottle 3: L-Malate dehydrogenase plus citrate synthase suspension, 2.2mL. Stable for > 2 years at 4°C.

Bottle 4: Acetyl-coenzyme A synthetase suspension (1.1 mL). Stable for> 2 years at 4°C.

REAGENT PREPARATION:

Preparation of R1:

Component Volume

bottle 1 (buffer) 5.50 mL

bottle 2 (NAD+/ATP/CoA) 2.20 mL (after adding 5.50 mL of H2O to bottle 2)

bottle 3 (L-MDH/CS) 0.44 mL (swirl to mix before use)

H2O 18.35 mL

Total volume 26.49 mL

Preparation of R2:

Component Volume

bottle 4 (ACS) 0.22 mL (swirl to mix before use)

H2O 1.60 mL

Total volume 1.82 mL

EXAMPLE METHOD:

R1: 0.290 mL

Sample: ~ 0.005 mL

R2: 0.020 mL

Reaction time: 15 min at either 20-25°C or 37°C

Wavelength: 340 nm

Prepared reagent stability: > 3 days when refrigerated

Calculation: endpoint

Reaction direction: increase

Linearity: up to 30 μg/mL of acetic acid in final reaction solution

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