A kind of cytochrome c detection method and kit
A detection method and cytochrome technology, applied in the direction of measuring devices, instruments, electrochemical variables of materials, etc., to achieve the effect of high sensitivity and convenient detection process
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Embodiment 1
[0035] Example 1: Preparation of Microbial Modified Screen Carbon Electrode
[0036] The microorganisms and the culture solution were inoculated in a 500ml conical flask at a ratio of 1:100, and cultured for 16 hours in a constant temperature shaker at a temperature of 30°C and a rotation speed of 180rpm. The culture solution was taken out, then centrifuged at 5000 g for 5 min, the precipitate was collected, washed 3 times with PBS (phosphate buffer, 0.02 M, pH 6), and the precipitate was resuspended with PBS. Add 7uL of microbial cell suspension dropwise to the surface of the working electrode of the screen-printed carbon electrode, and dry at room temperature (18-27° C.) for 60-120 min to bind the microbial cells to the surface of the working electrode.
[0037] In order to verify whether microbial cells modify the surface of the screen-printed carbon electrode, the present invention has carried out a scanning electron microscope characterization, and the results show that t...
Embodiment 2
[0041] Example 2: In situ detection of microbial cytochrome c
[0042]Schematic diagram of the method for in situ detection of cytochrome c as shown in figure 1 As shown, the specific operation steps are:
[0043] 1) Add 7uL of the microbial cell solution to be tested onto the working electrode surface of the screen-printed carbon electrode, and dry at room temperature (18-27°C) for 60-120min to bind the microbial cells to the surface of the working electrode to obtain a microbial-modified screen Printed carbon electrodes;
[0044] 2) Connect the prepared microbial-modified screen-printed carbon electrode to a CHI660D electrochemical workstation (Shanghai Chenhua Instrument Co., Ltd., China), select the detection mode as the time-current curve mode in the operating software of the electrochemical workstation, and set the voltage to -0.20V;
[0045] 3) Tetramethylbenzidine solution containing 0.2mg / ml tetramethylbenzidine, 10% v / v absolute ethanol, 0.7mMH 2 o 2 , 70% v / v (...
Embodiment 3
[0048] Example 3: In situ detection of microbial cytochrome c
[0049] Schematic diagram of the method for in situ detection of cytochrome c as shown in figure 1 As shown, the specific operation steps are:
[0050] 1) Add 7uL of the microbial cell solution to be tested onto the working electrode surface of the screen-printed carbon electrode, and dry at room temperature (18-27°C) for 60-120min to bind the microbial cells to the surface of the working electrode to obtain a microbial-modified screen Printed carbon electrodes;
[0051] 2) Connect the prepared microbial-modified screen-printed carbon electrode to a CHI660D electrochemical workstation (Shanghai Chenhua Instrument Co., Ltd., China), select the detection mode as the time-current curve mode in the operating software of the electrochemical workstation, and set the voltage to -0.20V;
[0052] 3) Tetramethylbenzidine solution containing 0.05mg / ml tetramethylbenzidine, 1% v / v absolute ethanol, 0.3mMH 2 o 2 , 30% v / v ...
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