Self-supplied consumables:
Microplate reader or visible light spectrophotometer (can measure the absorbance at 510 nm)
Thermostatic box, ice machine, low temperature centrifuge
96-well plate or microglass cuvette
Adjustable pipet gun and tip
deionized water
Homogenizer (if it is a tissue sample)

Reagent preparation:
Extraction Buffer: read-to-use type; Before use, equilibrate to room temperature; Store at 4 ° C.
Chromogen A: ready-to-use; Before use, equilibrate to room temperature; Store away from light at 4 ° C.
Chromogen B: ready-to-use type; Before use, equilibrate to room temperature; Store away from light at 4 ° C.
Chromogen C: ready-to-use type; Before use, equilibrate to room temperature; Store away from light at 4 ° C.
Standard: ready-to-use type; Before use, balance to room temperature; Store at 4 ° C.

Sample preparation:

Note: Fresh samples are recommended, and if experiments are not performed immediately, samples can be stored at -80° C 1 month.
1. Tissue samples: Weigh about 0.1 g sample, add 1 mL Extraction Buffer, homogenize in an ice bath, centrifuge at 10,000 rpm for 10 min at 4 ° C, take the supernatant and place it on ice for testing.
2. Serum (plasma) : plasma and serum can be directly used for determination. EDTA and citrate can not be used in plasma preparation, but other anticoagulants can be used.
3, urine: direct measurement.
Note: To measure protein concentration, use Item No. : abs9232  protein quantification kit (BCA method) was used to determine the protein concentration of samples.

Experimental procedures:
1. The microplate reader or visible light spectrophotometer was preheated for more than 30 min, and the wavelength was adjusted to 510 nm.
2. the table below for sample and response:

2-8℃, stored in the dark, valid for 12 months.