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Detection of tyrosine hydroxylase (TH) activity

JS Jiangbo Song
LL Lian Liu
ZL Zhiquan Li
TM Ting Mao
JZ Jianfei Zhang
LZ Lei Zhou
XC Xin Chen
YS Yunzhu Shang
TS Tao Sun
YL Yuxin Luo
YJ Yu Jiang
DT Duan Tan
XT Xiaoling Tong
FD Fangyin Dai
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The activity of insect TH was determined using a double-antibody sandwich method with an insect TH enzyme-linked immunosorbent assay kit (Jiangsu Enzyme Immune Industrial Co., Ltd., Yancheng, China). The sample was ground into powder and fully dissolved with PBS to determine the protein concentration. Fifty-microliter standards were added in the control group, while 40-μL sample diluent and 10-μL sample were added in the treatment group, mixed well, and the plate was sealed. Standards and sample mixtures were incubated for 30 min at 37 °C and washed five times. Then, 50-μL enzyme-linked immunosorbent assay reagent was added. Standards and sample mixtures were incubated for 30 min at 37 °C after washing five times. The following colour development step was performed according to the kit instructions. The absorbance was measured at 450 nm within 15 min after termination of the reaction. The protein activity of the sample was calculated according to the standard curve.

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