Products
Monoamine Oxidase (MAO) Assay Kit(Colorimetric)
- Catalog No.
K059
- Size
50T/100T
- Storage
The kit should be stored at 2-8℃ for one year.
- Intended use
This kit can be used to measure monoamine oxidase (MAO) activity in serum, plasma and animal tissue samples.
- Sensitivity
16 U/L
- Detection range
16-641 U/L
- Introduction
MAO can catalysis 4-dimethylambenzylamine to produce p-dimethylaminobenzaldehyde. p-Dimethylaminobenzaldehyde has a characteristic absorption peak at 355nm. The activity of MAO can be calculated indirectly by analyzing the production of p-dimethylaminobenzaldehyde.
- Kit Components
Item
50T
100T
Reagent 1
30mL
60mL
Reagent 2
60mL
60mL*2
Reagent 3
60mL
60mL*2
Reagent 4
3mL
5mL
- Materials prepared by users
Microplate Reader(355 nm)
- Reagent preparation
1.The preparation of Reagent 1 working solution:
Mix Reagent 1 with double distilled water fully at a ratio of 1:1. The prepared solution can be stored at 2-8℃ for 1 month.
2.The preparation of Reagent 3 working solution:
Mix reagent 3 with double distilled water fully at a ratio of 1:1.The prepared solution can be stored at 2-8℃ for 1 month.
- Sample preparation
1.The preparation of Reagent 1 working solution:
Mix Reagent 1 with double distilled water fully at a ratio of 1:1. The prepared solution can be stored at 2-8℃ for 1 month.
2.The preparation of Reagent 3 working solution:
Mix reagent 3 with double distilled water fully at a ratio of 1:1.The prepared solution can be stored at 2-8℃ for 1 month.
Sample preparation
1.Serum or plasma: detect directly.
2.10% tissue homogenate: Accurately weigh the tissue 0.1-0.5g, then add 9 times the volume of pre-cooled Reagent 1 working solution according to the ratio of Weight (g): Volume (mL)=1:9. The sample is mechanically homogenized for 90s in an ice water bath. Centrifuge at 1000 g for 10 min at 4°C, then take the supernatant(determine the protein concentration of supernatant (K001) before centrifugation) and centrifuge at 10000 g at 4°C for 30 min, discard the supernatant and retain the precipitation. Add 1 mL of pre-cooled Reagent 2 and mix fully, centrifuge at 16,000 g at 4°C for 40 min, discard the supematant and retain the precipitation. Finally, add 1 mL of pre-cooled Reagent 3 working solution, mix fully, and store it on ice for detection.
Note: Reagent 1 working solution, Reagent 2, Reagent 3 working solution need to be pre-cooled for 30min in advance.
- Assay Procedure
1.Reagent 3 working solution and Reagent 4 need to be pre-heated at 37°C for 30 min in advance.
2.Add reagents according to the table below:
reagents
Sample tube(μL)
Sample
25
Reagent 3 working solution
150
Reagent 4
25
3.Vibrate on Microplate Readerr for five seconds, measure the OD value of samples at 355 nm, recorded as A1, and then incubate accurately at 37℃ for 30 min, measure the OD values of each samples again, recorded as A2.
- Calculation
1.Serum or plasma sample:
Definition: the amount of enzyme in 1 L of serum (plasma) that catalyze the substrate to produce 1 nmol p-dimethylaminobenzaldehyde at 37 C for 1 min is defined as 1 unit.
2.Tissue sample:
Definition: the amount of enzyme in 1 g of tissue protein that catalyze the substrate to produce 1 nmol p-dimethylaminobenzaldehyde at 37℃ for 1 min is defined as 1 unit.
Note:
T: the time of incubation in the reaction, 30min
ε: the molar extinction coefficient of p-dimethylaminobenzaldehyde, 2.77×10-4L/nmol.cm)
d: the optical path of cuvette, 0.6cm.
V1: the total volume of reaction, 200μl.
V2: the volume of sample, 25μl.
Cpr: The concentration of protein in sample, gprot/L.