Application

This kit can be used for detection of total amino acid (T-AA) content in samples, such as tissue, urine, serum (plasma) and other samples. 

Detection significance

Liver and kidney are the main organs of amino acid metabolism, therefore the change of amino acid in urine can reflect the pathological and physiological state of liver and kidney. In addition, amino acid can reflect the condition of burns, diabetes, typhoid and other diseases.

Detection principle

Cu2+ can react with all kinds of amino acids to form blue-green complex. The depth of color is proportional to the content of total amino acids at 650 nm. The content of total amino acid can be calculated by measuring the OD value.   

Experimental instrument

Test tube, Micropipettor, Vortex mixer, Centrifuge, Water bath, Spectrophotometer (650 nm)

Operation steps

1. For urine sample

Mix fully, then centrifuge at 3500 rpm for 10 min. Set the spectrophotometer to zero with double-distilled water and take the supernatant to measure the OD values of each tube at 650 nm with 1 cm optical path cuvette. 

2. Foe serum (plasma) sample

Mix fully, then centrifuge at 3500 rpm for 10 min. Set the spectrophotometer to zero with double-distilled water and take the supernatant to measure the OD values of each tube at 650 nm with 1 cm optical path cuvette.

3. For tissue sample

(1) Sample pretreatment:

Weigh the tissue accurately, then add normal saline at the ratio of Weight (g): Volume (mL) = 1:9. Make the mechanical homogenization in ice water bath to prepare 10% tissue homogenate. Centrifuge the tissue homogenate at 3500 rpm for 10 min and take the supernatant for detection. Meanwhile, determine the protein concentration of supernatant (E-BC-K318, E-BC-K168, E-BC-K165)

(2) Operation table:

Mix fully, then centrifuge at 3500 rpm for 10 min. Set the spectrophotometer to zero with double-distilled water and take the supernatant to measure the OD values of each tube at 650 nm with 1 cm optical path cuvette.

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. It must be slow when adding Reagent 2, and the suspension must be fully dissolved as possible. The reaction solution should be light-blue transparent solution.

6. The Reagent 3 is corrosive, it should be careful when preparing.

7. The test tubes in this experiment must be boiled with hot soapy water, then wash with running water. Otherwise, measuring the activity of other enzymes with these tubes will affect the results. It is recommended to use disposable plastic tubes.