CK-MB NAC activated Macro

Pipette directly into the reconstituted working reagent

Humazym M-Test Sample 100 µl
Immunoinhibition Method for CK-MB Mix and transfer the solution to a cuvette. Incubate at the desired
Creatine Kinase (EC 2.7.3.2) temperature for 10 minutes. Read the absorbance A 1. Exactly after 5
min read the absorbance A2.
Package Sizes
[REF] 12008 20 x 3 ml Complete test kit Semi-micro
[IVD] Pipette into cuvettes
Sample 40 µl
Method
Working reagent 1000 µl
HumaZym CK-MB is based on an enzymatic CK determination
accompanied by an immunoinhibition method1,2. An antibody is Mix and incubate at the desired temperature for 10 minutes.
incorporated into the reagent which will bind specifically to the M- Read the absorbance A1. Exactly after 5 min read the absorbance A2.
subunit, inhibiting the enzymatic activity of that subunit. Thus only the
remaining activity of the B-subunit is measured. Due to negligible Calculation
concentrations of CK-BB in the circulation, the remaining activity, Calculate the absorbance change per minute ( A/min) according to:
multiplied by the factor 2, represents the activity of the CK-MB A/min = (A2-A1) : 5.
isoenzyme.
The CK-MB activity is calculated using the following factors:
Principle CK-MB activity (U/l)
CK
Semi-micro Macro
Creatine phosphate + ADP Creatine + ATP
Hg 334 nm 8414 x A/min 10032 x A/min
340 nm 8254 x A/min 9842 x A/min
HK
Hg 365 nm 14858 x A/min 17716 x A/min
ATP + D-glucose ADP + D-glucose-6-
Conversion factor of traditional units (U/l) in SI-units (kat/l)
phosphate (G6P)
1 U/l = 16.67 x 10-3 µkat/l
1 µkat/l = 60 U/l
G6P-DH
G6P + NADP 6-phospho-D-glucono¬ Performance Characteristics
lactone + NADPH + H + Linearity: The antibody inhibits the activity of M-subunits up to 2000 U/l
of CK MM.
Typical performance date can be found in the Verification Report,
Contents, Reagent Composition in the Test accessible via
[BUF] 1 x 60 ml buffer solution www.human.de/data/gb/vr/en-ckmb.pdf or
Imidazole buffer (pH 6.7) 0.10 mol/l www.human-de.com/data/gb/vr/en-ckmb.pdf
Glucose 20 mmol/l
Mg-acetate 10 mmol/l Reference Range, Myocardial Infarction (MI)
EDTA 2.00 mmol/l The likelihood of myocardial damage is high if the following 3 criteria are
met:
[ENZ] 20 x 3 ml enzyme/antibody reagent (lyoph.)
ADP 2.00 mmol/l 25°C3 30°C4 37°C
AMP 5.00 mmol/l 1. Total CK
Diadenosine pentaphosphate 10 mmol/l men > 80 U/l > 130 U/l > 195 U/l
NADP 2.00 mmol/l
women > 70 U/l > 110 U/l > 170 U/l
HK > 2.50 U/ml
G6P-DH > 1.50 U/ml 2. CK-MB > 10 U/l > 16 U/l > 25 U/l
N-Acetylcysteine 20 mmol/l 3. CK-MB activity ranging between 6% and 25% of the total CK activity
Creatine phosphate 30 mmol/l
Antibody to CK-M subunit (goat) Quality Control
All control sera with CK-MB values determined by this method can be
Reagent Preparation and Stability employed. Only control sera with human CK can be used.
Reconstitute one vial [ENZ] with exactly 3 ml [BUF]. Swirl gently and
incubate for 5 min. at room temperature prior to use. Automation
This kit is designed for manual use. For use on automates we recommend
The reagents are stable up to the stated expiry date when stored at
[REF] 12118. Proposals to apply this reagents on analysers are available
2...8°C.
on request. Each laboratory has to validate the application in its own
The reconstituted working reagent is stable for 5 days at 2...8°C. responsibility.
Specimen Note
Serum, heparinised plasma or EDTA plasma. [BUF] contains sodium azide (< 0.095%) as stabiliser. Avoid contact with
Loss of activity within 1 day at 2...8°C: < 10%. the skin and mucous membranes.

Assay References
Wavelength: Hg 334 nm, 340 nm, Hg 365 nm 1. Würzburg, U. et al., Klin. Wschr. 54, 357 (1976)
Optical path: 1 cm 2. Würzburg, U. et al., J. Clin. Chem. Clin. Biochem. 15, 131 (1977)
Temperature: 25°C, 30°C or 37°C 3. Stein, W., medwelt 36, 572 (1985)
Measurement: against air (increasing absorbance) 4. Szasz, G. and Busch, E.W., Abstract presented at 3 rd Eur. Congr. Clin.
Chem., Brighton/UK, 3-8 (1979)
Procedure
Prior to determining CK-MB activity it is recommended to measure total
CK activity using the CK-NAC activated method (CK NAC liquiUV, [REF]
EN-CKMB INF 1200801 GB 03-2009-14 |
12015, HumaZym M-Test, [REF] 12005) in order to ensure correct
diagnostic interpretation of the test results.
Warm reagents and cuvettes to the desired temperature. Temperature
must be kept constant (± 0.5°C) for the duration of the test.

Human Gesellschaft für Biochemica und Diagnostica mbH

Max-Planck-Ring 21 · 65205 Wiesbaden · Germany
Telefon +49 6122-9988-0 · Telefax +49 6122-9988-100 · e-Mail human@human.de