Application

This kit can be used to detect the maltase activity in animal tissue and other samples. 

Detection significance

Maltase in the small intestine is a key enzyme of the carbohydrate absorption in the digestive tract. Maltase is an enzyme located in on the brush border of the small intestine that breaks down the disaccharide maltose. Maltase catalyzes the hydrolysis of maltose to the simple sugar glucose. This enzyme is found in plants, bacteria, and yeast. If the organism lacks lactase or other Maltase, will lead to the disorder of carbohydrate digestion and absorption of food, the Undigested and Unabsorbed carbohydrates get into the large intestine, then be decomposed by bacteria into CO2, H2 and so on, give rise to diarrhea and other symptoms.

Detection principle

Maltase catalyze the corresponding substrate to produce monosaccharide. Monosaccharide produce hydrogen peroxide under the action of oxidase. Hydrogen peroxide react with chromogenic agent to form red product. The activity of maltase can be calculated by detection the optical density with spectrophotometer at 505 nm.

Experimental instrument

Test tube, Micropipettor, Vortex mixer,Centrifuge, 37℃ incubator, Spectrophotometer (505 nm) or Microplate reader (505 nm)

The preparation of tissue sample

10% tissue homogenate sample: Accurately weigh the tissue sample, add 9 times the volume of PBS (0.01 M, pH7~7.4) according to the ratio of Weight (g): Volume (mL) =1:9. Mechanical homogenate the sample in ice water bath. Centrifuge at 3500 g for 10 min, then take the supernatant and preserve it on ice for detection. Meanwhile, determine theprotein concentration of supernatant (E-BC-K318, E-BC-K168, E-BC-K165).

Operation steps

1. Enzymatic reaction

Substrate solution (μL) 

2. Chromogenic reaction

1) Measured by microplate reader

Supernatant (μL)  

Chromogenic agent (μL) 

2) Measured by spectrophotometry

Supernatant (μL)  

Chromogenic agent (μL) 

[Note] The optimum sampling concentration should be determined by pre-experiment before batch experiment,The (ODsample-ODcontrol) value should be closed to ODstandard. 

Notes

1. The kit is for scientific research only.

2. Instructions should be followed strictly, changes of operation may result in unreliable results.

3. The validity of kit is 6 months.

4. Do not use components from different batches of kit.

5. If the maltase activity is calculated by protein concentration, the protein concentration of the sample needs to be determined separately (E-BC-K318, E-BC-K168, E-BC-K165).